Journal articles: 'Cryopreservation'

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Koung Ngeun, Sai, Miki Shimizu, and Masahiro Kaneda. "Characterization of Rabbit Mesenchymal Stem/Stromal Cells after Cryopreservation." Biology 12, no. 10 (October 7, 2023): 1312. http://dx.doi.org/10.3390/biology12101312.

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Adipose tissues (ADPs) are an alternative source for mesenchymal stem/stromal cells (MSCs), given that conventional bone marrow (BM) collection is painful and yields limited cell numbers. As the need for easily accessible MSCs grows, cryopreservation’s role in regenerative medicine is becoming increasingly vital. However, limited research exists on the characteristics and functional properties of rabbit-derived MSCs from various anatomical sources before and after cryopreservation. We examined the effects of cryopreservation using Bambanker. We found that cryopreservation did not adversely affect the morphology, viability, and adipogenic or chondrogenic differentiation abilities of ADP MSCs or BM MSCs. However, there was a notable drop in the proliferation rate and osteogenic differentiation capability of BM MSCs post-cryopreservation. Additionally, after cryopreservation, the surface marker gene expression of CD90 was not evident in ADP MSCs. As for markers, ADIPOQ can serve as an adipogenic marker for ADP MSCs. ACAN and CNMD can act as chondrogenic markers, but these two markers are not as effective post-cryopreservation on ADP MSCs, and osteogenic markers could not be validated. The study highlights that compared to BM MSCs, ADP MSCs retained a higher viability, proliferation rate, and differentiation potential after cryopreservation. As such, in clinical MSC use, we must consider changes in post-cryopreservation cell functions.

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Takeuchi, Hiroki, Mikiko Nishioka, Tadashi Maezawa, Yuko Kitano, Kento Terada-Yoshikawa, Ryota Tachibana, Manabu Kato, et al. "Carboxylated Poly-l-Lysine as a Macromolecular Cryoprotective Agent Enables the Development of Defined and Xeno-Free Human Sperm Cryopreservation Reagents." Cells 10, no. 6 (June 8, 2021): 1435. http://dx.doi.org/10.3390/cells10061435.

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In human sperm cryopreservation, test yolk buffer and human serum albumin have been used as permeating macromolecular-weight cryoprotectants. In clinical reproductive medicine, human serum albumin is frequently used because of low risks of zoonoses and allergic reactions. However, the risk of allogeneic infectious diseases exists, and the supply may be unstable because human serum albumin is derived from human blood. Therefore, the development of xeno-free human sperm cryopreservative reagents that could overcome the aforementioned problems is warranted. We succeeded in developing a new xeno-free and defined sperm cryopreservation reagent containing glycerol, carboxylated poly-l-lysine, and raffinose. The cryopreservation reagent was not significantly different in terms of sperm motility, viability, and DNA fragmentation and was comparable in performance to a commercial cryopreservation reagent containing human serum albumin. Moreover, the addition of saccharides was essential for its long-term storage. These results may help elucidate the unknown function of macromolecular-weight permeating cryoprotective agents.

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Adams, R. Mark, Mary Wang, Ana Maria Crane, Bridgette Brown, Gretchen J. Darlington, and Fred D. Ledley. "Effective Cryopreservation and Long-Term Storage of Primary Human Hepatocytes with Recovery of Viability, Differentiation, and Replicative Potential." Cell Transplantation 4, no. 6 (November 1995): 579–86. http://dx.doi.org/10.1177/096368979500400607.

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Despite reports of successful cryopreservation of primary human hepatocytes, existing methods do not produce sufficient recovery of viable cells to meet the needs of basic research or clinical trials of hepatocellular transplantation. We now describe a protocol for efficient cryopreservation of primary human hepatocytes using University of Wisconsin (UW) solution, fetal bovine serum, and dimethyl sulfoxide (DMSO). This method provides >90% viability of differentiated, primary human hepatocytes 8 mo after cryopreservation as measured by trypan blue exclusion, preserves hepatocyte morphology, liver-specific gene expression α1 antitrypsin), and replication. The effectiveness of UW solution as a cryopreservative agent suggests that metabolic as well as ultrastructural factors may be important in the effective cryopreservation of primary human hepatocytes. The present method represents an effective protocol for cryopreserving differentiated primary human hepatocytes for research. This method may allow characterization and banking of human hepatocytes for clinical applications, including hepatocellular transplantation and hepatic assist devices.

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Crowley, Conor A., William P. W. Smith, K. T. Matthew Seah, Soo-Keat Lim, and Wasim S. Khan. "Cryopreservation of Human Adipose Tissues and Adipose-Derived Stem Cells with DMSO and/or Trehalose: A Systematic Review." Cells 10, no. 7 (July 20, 2021): 1837. http://dx.doi.org/10.3390/cells10071837.

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Adipose tissue senescence is implicated as a major player in obesity- and ageing-related disorders. There is a growing body of research studying relevant mechanisms in age-related diseases, as well as the use of adipose-derived stem cells in regenerative medicine. The cell banking of tissue by utilising cryopreservation would allow for much greater flexibility of use. Dimethyl sulfoxide (DMSO) is the most commonly used cryopreservative agent but is toxic to cells. Trehalose is a sugar synthesised by lower organisms to withstand extreme cold and drought that has been trialled as a cryopreservative agent. To examine the efficacy of trehalose in the cryopreservation of human adipose tissue, we conducted a systematic review of studies that used trehalose for the cryopreservation of human adipose tissues and adipose-derived stem cells. Thirteen articles, including fourteen studies, were included in the final review. All seven studies that examined DMSO and trehalose showed that they could be combined effectively to cryopreserve adipocytes. Although studies that compared nonpermeable trehalose with DMSO found trehalose to be inferior, studies that devised methods to deliver nonpermeable trehalose into the cell found it comparable to DMSO. Trehalose is only comparable to DMSO when methods are devised to introduce it into the cell. There is some evidence to support using trehalose instead of using no cryopreservative agent.

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Khan, Muhammad Shoaib, Mohd Basyaruddin Abdul Rahman, Adamu Abdul Abubakar, Shakeeb Ullah, Humaira A. Khan, Farheen Bhatti, Nayab Batool Rizvi, et al. "AN OVERVIEW OF TISSUE CRYOPRESERVATION'S CONTRIBUTION TO ORGAN CRYOBIOLOGY." Health Sciences Journal 2, no. 1 (December 31, 2023): 8–15. http://dx.doi.org/10.59365/hsj.2(1).2023.54.

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Background: Scientists and physicians have long faced challenges in their quest to identify the most effective methods for cryopreservation of living tissues and organs. While cryopreservation techniques have been used since the 1800s, they are still used for tissue preservation for a comparatively limited time. More advancements in biotechnology for tissue and organ preservation have been made possible by the availability of commercially available liquefied gases. Typical issues such as sub-zero or cell damage and re-crystallization, which ultimately results in tissue loss, impede the usefulness of cryopreservation in the field of medical sciences. However, modern cryobiology advances have provided many alternatives for chilling tissues, organs, gametes, and embryos. As tissue sensitivity to ice is more understood, safer freezing methods are used. Methods: This review discusses how tissue cryopreservation advances biotechnology. The effective banking and transplanting of living tissues and organs using cryopreservation will be a crucial and noteworthy development in the fields of biotechnology and medicine in the future. One of the most crucial elements that will enable future cryobiologists and biotechnology researchers to make enormous strides is the preservation and storage of living tissues from the point of donor removal through transfer and transplantation. Future cryobiologists will find this ancient tale fascinating as it unfolds, thanks to the discovery of anti-freezing proteins and peptides in cryopreservation's active zones.

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Lederman, Lynne. "Cryopreservation." BioTechniques 45, no. 6 (December 2008): 619–21. http://dx.doi.org/10.2144/000112986.

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Baust, John G., Dayong Gao, and John M. Baust. "Cryopreservation." Organogenesis 5, no. 3 (July 2009): 90–96. http://dx.doi.org/10.4161/org.5.3.10021.

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Trounson, A. O. "Cryopreservation." British Medical Bulletin 46, no. 3 (1990): 695–708. http://dx.doi.org/10.1093/oxfordjournals.bmb.a072425.

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Pe, Phyo Phyo Win, Aung Htay Naing, Chang Kil Kim, and Kyeung Il Park. "Antifreeze Protein Improves the Cryopreservation Efficiency of Hosta capitata by Regulating the Genes Involved in the Low-Temperature Tolerance Mechanism." Horticulturae 7, no. 4 (April 15, 2021): 82. http://dx.doi.org/10.3390/horticulturae7040082.

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In this study, whether the addition of antifreeze protein (AFP) to a cryopreservative solution (plant vitrification solution 2 (PVS2)) is more effective in reducing freezing injuries in Hosta capitata than PVS2 alone at different cold exposure times (6, 24, and 48 h) is investigated. The upregulation of C-repeat binding factor 1 (CBF1) and dehydrin 1 (DHN1) in response to low temperature was observed in shoots. Shoots treated with distilled water (dH2O) strongly triggered gene expression 6 h after cold exposure, which was higher than those expressed in PVS2 and PVS2+AFP. However, 24 h after cold exposure, gene expressions detected in dH2O and PVS2 treatments were similar and higher than PVS2 + AFP. The expression was highest in PVS2+AFP when the exposure time was extended to 48 h. Similarly, nitric reductase activities 1 and 2 (Nia1 and Nia2) genes, which are responsible for nitric oxide production, were also upregulated in low-temperature-treated shoots, as observed for CBF1 and DHN1 expression patterns during cold exposure periods. Based on the gene expression patterns, shoots treated with PVS2+AFP were more likely to resist cold stress, which was also associated with the higher cryopreservation efficiency of PVS2+AFP compared to PVS2 alone. This finding suggests that the improvement of cryopreservation efficiency by AFP could be due to the transcriptional regulation of CBF1, DHN1, Nia1, and Nia2, which might reduce freezing injuries during cryopreservation. Thus, AFP could be potentially used as a cryoprotectant in the cryopreservation of rare and commercially important plant germplasm.

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Jovičić, Marija, Eva Chmelíková, and Markéta Sedmíková. "Cryopreservation of boar semen." Czech Journal of Animal Science 65, No. 4 (April 30, 2020): 115–23. http://dx.doi.org/10.17221/47/2020-cjas.

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Sperm cryopreservation is the best technology for long-term storage of the semen. However, the damage of boar spermatozoa by cryopreservation is more severe than in other animal species and a standardized freezing protocol for efficient cryopreservation has not been established yet. Semen quality and freezability vary greatly between breeds as well as between individual boars and even the season. Boar spermatozoa are sensitive to low temperatures; they sustain damage and a high rate of mortality and freezing/thawing the boar semen may strongly impair the sperm function and decrease the semen quality. The freezability of boar semen can be influenced by a cryopreservation procedure, and also by using various additives to freezing and thawing extenders such as antioxidants. In order to obtain acceptable results after thawing the boar semen, it is necessary to combine an optimal amount of additives (glycerol, egg yolk, sugars, antioxidants), cooling and warming velocities.

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Baran, S., and C. Ware. "83ANALYSIS OF RAPID COOLING V. SLOW COOLING COMBINED WITH ICE CRYSTAL SEEDING FOR CRYOPRESERVATION OF PRIMATE EMBRYONIC STEM CELLS." Reproduction, Fertility and Development 16, no. 2 (2004): 163. http://dx.doi.org/10.1071/rdv16n1ab83.

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Primate embryonic stem (ES) cells have the ability to self-renew indefinitely while maintaining the ability to differentiate. This unique property allows scientists to study the factors necessary for stem cell self-renewal and differentiation in vitro that reflect in vivo processes. Work with primate ES cells is handicapped by the poor survival (1–5%) of rhesus and human ES cells following standard tissue culture methods of rapid cryopreservation. The purpose of this study was to compare and contrast two cryopreservation techniques, slow cooling combined with ice crystal seeding commonly used for mammalian embryos v. rapid cooling commonly used for tissue culture, to find a method for efficient primate ES cell cryopreservation. A combination of trials was run to compare dimethyl sulfoxide (DMSO) v. ethylene glycol as a cryoprotectant, a cooling rate of 0.3°C per minute following ice crystal seeding at −7°C v. placement at −80°C with no seeding, and rapid thaw with step-wise cryoprotectant removal v. one-step sucrose cryoprotectant removal. Cell survival was assessed through a combination of cell surface markers, alkaline phosphatase staining and morphology to look for undifferentiated cells and quantitate survival. All cryopreservations were performed with the same cell density. The survival of the cells with slow embryo-style cooling in DMSO with a step-wise cryoprotectant removal was 64.0% v. 12.8% with rapid cooling.

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Cedars, Marcelle. "Embryo Cryopreservation." Seminars in Reproductive Medicine 16, no. 03 (September 1998): 183–95. http://dx.doi.org/10.1055/s-2007-1016277.

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Fabbri, R., E. Porcu, T. Marsella, M. R. Primavera, R. Seracchioli, P. M. Ciotti, O. Magrini, S. Venturoli, and C. Flamigni. "Oocyte cryopreservation." Human Reproduction 13, suppl 4 (December 1, 1998): 98–108. http://dx.doi.org/10.1093/humrep/13.suppl_4.98.

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Trounson, AO, and JM Shaw. "Embryo cryopreservation." Reproductive Medicine Review 1, no. 2 (July 1992): 179–88. http://dx.doi.org/10.1017/s0962279900000521.

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Embryo cryopreservation is a well established technique and is used widely for embryo banking of genetically valuable strains of mice, the transport and proliferation of farm animals and as a valuable adjunct to human in vitro fertilization (IVF). The range of methods presently used to cryopreserve human embryos has been recently reviewed. This article examines the results obtained by the established freezing techniques and developments in the new rapid cooling methods. There is a dramatic contrast in the simplicity, ease and cost between these new rapid techniques and the conventional slow cooling or equilibrium freezing methods and it is likely that the rapid freezing will replace conventional freezing by slow cooling which is presently in widespread use in IVF clinics.

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Feichtinger, W. "Cryopreservation techniques." Fertility and Sterility 79, no. 5 (May 2003): 1255. http://dx.doi.org/10.1016/s0015-0282(02)04954-3.

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Albani, E., J. Barbieri, P. V. Novara, A. Smeraldi, G. Scaravelli, and P. E. Levi Setti. "Oocyte Cryopreservation." Placenta 29 (October 2008): 143–46. http://dx.doi.org/10.1016/j.placenta.2008.08.002.

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Vendrame, Wagner, Ricardo Tadeu de Faria, Mauren Sorace, and Sandra Aparecida Sahyun. "Orchid cryopreservation." Ciência e Agrotecnologia 38, no. 3 (June 2014): 213–29. http://dx.doi.org/10.1590/s1413-70542014000300001.

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Orchids are lush and highly valuable plants due to their diversity and the beauty of their flowers, which increases their commercialization. The family Orchidaceae comprises approximately 35,000 species, distributed among more than 1,000 distinct genera and 100,000 hybrids, totaling approximately 8% to 10% of all flowering plants. With the advance of agriculture and the constant destruction of their natural habitat, orchid species are collected in an indiscriminate manner by collectors and vendors, and this extractive activity threatens many species with extinction, drastically reducing their genetic variability in nature. Therefore, it is essential to seek alternatives that make the preservation of such species feasible using techniques with low maintenance costs that provide greater storage time and that enable good phytosanitary conditions for the plant material for commercial use. Cryopreservation involves the conservation of biological materials at ultra-low temperatures, generally in liquid nitrogen at -196 ºC or in its vapor phase at -150 ºC. This is the only technique currently available for the long-term preservation of the germplasm of plant species that are vegetatively propagated or that have unviable, recalcitrant or intermediate seeds. The objective of this bibliographic review is to report on the importance, methods and application of cryopreservation for orchids. According to the studies reviewed, this is an incipient, developing and relevant field that generates a lot of discussion and requires further research relative to the type of treatment to use for cryopreservation and the methodology to be applied according to the species. The types of methods that are used for cryopreservation and the large variation in the responses of orchids to the cryopreservation methods observed in this study emphasize the need for the development of more appropriate protocols for the preservation of orchids.

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Kuwayama, Masashige. "Oocyte Cryopreservation." Journal of Mammalian Ova Research 24, no. 1 (April 2007): 2–7. http://dx.doi.org/10.1274/jmor.24.2.

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JAIN, J., and R. PAULSON. "Oocyte cryopreservation." Fertility and Sterility 86, no. 4 (October 2006): 1037–46. http://dx.doi.org/10.1016/j.fertnstert.2006.07.1478.

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Zhang, Chaocan, Youliang Zhou, Li Zhang, Lili Wu, Yanjun Chen, Dong Xie, and Wanyu Chen. "Hydrogel Cryopreservation System: An Effective Method for Cell Storage." International Journal of Molecular Sciences 19, no. 11 (October 25, 2018): 3330. http://dx.doi.org/10.3390/ijms19113330.

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At present, living cells are widely used in cell transplantation and tissue engineering. Many efforts have been made aiming towards the use of a large number of living cells with high activity and integrated functionality. Currently, cryopreservation has become well-established and is effective for the long-term storage of cells. However, it is still a major challenge to inhibit cell damage, such as from solution injury, ice injury, recrystallization and osmotic injury during the thawing process, and the cytotoxicity of cryoprotectants. Hence, this review focused on different novel gel cryopreservation systems. Natural polymer hydrogel cryopreservation, the synthetic polymer hydrogel cryopreservation system and the supramolecular hydrogel cryopreservation system were presented, respectively. Due to the unique three-dimensional network structure of the hydrogel, these hydrogel cryopreservation systems have the advantages of excellent biocompatibility for natural polymer hydrogel cryopreservation systems, designability for synthetic polymer hydrogel cryopreservation systems, and versatility for supramolecular hydrogel cryopreservation systems. To some extent, the different hydrogel cryopreservation methods can confine ice crystal growth and decrease the change rates of osmotic shock in cell encapsulation systems. It is notable that the cryopreservation of complex cells and tissues is demanded in future clinical research and therapy, and depends on the linkage of different methods.

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C. S., Sushrutha, Sandhya K., Savitha Karlwad, and Elango E. M. "Assessment of albumin and aspartate levels as a simple indicator of the efficacy of cryopreservation." International Journal of Scientific Reports 7, no. 2 (January 22, 2021): 110. http://dx.doi.org/10.18203/issn.2454-2156.intjscirep20210094.

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Background: The process of hepatocytes cryopreservation is standardised by most of the laboratories. However there is a variation with respect to the Protocols, media and equipments used amongst the laboratories. Similarly, the tests available to evaluate the efficacy also varies. They are expensive and sometimes might not measure the parameter required for a particular research study. Hence we propose a methodology to study the few basic parameters like cell viability, synthetic function of the cell and cell stability. We have also used a simple percentile calculation to know the efficacy of cryopreservation. This shall help in functional validation of the cell after cryopreservation. The same can also be used to compare the quality of hepatocytes between different batches. The objective of the study was to characterisation of the cells to determine the efficacy of cryopreservation.Methods: Two step collagen isolation method was used to isolate the hepatocytes. Initial cell viability was calculated. A sample of cells were taken for characterisation and the remaining cells cryopreserved. The sample cells were divided into two batches one for pre cryopreservation culture and the other for post cryopreservation. The pre cryopreservation culture was done on monolayer using collagen coated 6 well plate. The other sample was placed in the cryovials for cryopreservation for 1week. After 1 week the cryopreserved cells were thawed and the post cryopreservation viability calculated, followed by post cryopreservation culture. During the process of culture (both pre cryopreservation and post cryopreservation) for 5days Albumin was measured daily and average calculated, peak Aspartate (AST) at 24 hours was recorded. The percentile difference of the obtained values between the pre cryopreservation and post cryopreservation culture was calculated.Results: A total of 12 specimen were enrolled for the study. The mean pre cryopreservation viability of the cells was 66.58%. The post cryopreservation, viability of the cells was 36.43%. The mean difference was -30.170%. The pre cryopreservation albumin values had a mean of 150ng/ml. The post cryopreservation albumin values had a mean of 135.83ng/ml. The mean difference was -14.170ng/ml. The pre cryopreservation peak Aspartate values had a mean of 234.17 IU/ml. The post cryopreservation peak aspartate values had a mean of 230 IU/ml. The mean difference was -4.176 IU/ml.Conclusions: This simple method can validate the cells after cryopreservation by measurement of cell viability, synthetic function of the cell and cell stability.

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Javed, Murid, Navid Esfandiari, Jason Swain, and Essam Michael. "Human Oocyte Cryopreservation - An Emerging ART Technique: Are We Heading in the Right Direction?" Journal of Reproductive and Stem Cell Biotechnology 2, no. 2 (December 2011): 109–20. http://dx.doi.org/10.1177/205891581100200205.

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Oocyte cryopreservation is a promising adjunct to human assisted reproductive technology. Slow rate freezing has been the cryopreservation standard for storage of sperm, embryos and, subsequently, oocytes. However, earlier concerns regarding damage to the meiotic spindle, loss of cortical granules and the low success rates as compared to the relative success of embryo cryopreservation caused little interest until the 1990s. Interest increased when many studies indicated that acceptable oocyte survival, in vitro fertilization, normal embryos and adequate blastocyst development can be achieved with oocyte cryopreservation. Recently introduced oocyte vitrification techniques are proving to be more efficient. Survival rates are close to 100% and developmental rates are similar to those achieved with fresh oocytes. This efficiency opens the way to the widespread application of the technique in various medical, legal, and social situations, even to replace embryo cryopreservation with the oocyte cryopreservation. Oocyte vitrification has dominated slow freezing to such an extent that it may soon become the exclusive cryopreservation choice, especially considering that potential disease transmission problems commonly associated with vitrification due to direct exposure of oocytes to liquid nitrogen can be eliminated by using the proper techniques and devices. Furthermore, cryopreservation of immature oocytes, ovarian follicles, ovarian tissue and whole ovary are other emerging technologies. Oocyte cryopreservation has tremendous opportunity for preserving fertility in cancer patients, for those who may not have sperm following oocyte retrieval and for those women who wish to delay their motherhood. The purpose of this article is to review the history of oocyte cryopreservation, its applications, current cryopreservation techniques and future trends for fertility cryopreservation, to determine if oocyte cryopreservation is proceeding in the right direction.

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HOULE, DAVID, ALEXEY S. KONDRASHOV, LEV YU YAMPOLSKY, SHANNON CALDWELL, and PETER L. STEPONKUS. "The effect of cryopreservation on the lethal mutation rate in Drosophila melanogaster." Genetical Research 69, no. 3 (June 1997): 209–13. http://dx.doi.org/10.1017/s0016672397002760.

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Although cryopreservation is routinely used for the storage of a range of biological organisms, few studies have been conducted to determine whether cryopreservation increases the frequency of mutation. A procedure for the cryopreservation of Drosophila melanogaster embryos has recently been developed. Cryopreservation of D. melanogaster is of special interest to geneticists and evolutionary biologists because it would make it possible to assay control and experimental populations simultaneously during long-term studies. Before cryopreserved embryos can be used for such studies, it is first necessary to show that cryopreservation is not mutagenic. We tested for mutagenic effects of cryopreservation in D. melanogaster embryos with an X-linked, recessive lethal assay. The mutation rates of cryopreserved and control flies were not significantly different. We can be 95% certain that cryopreservation does not increase mutation by a factor greater than 2·39. This is the first quantitative estimate of the mutagenic effect of cryopreservation on the germ line of a metazoan. The results are reassuring when considering the genetic impact of cryopreservation on mammalian gametes and embryos.

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Jaing, Tang-Her, Shih-Hsiang Chen, Yu-Chuan Wen, Tsung-Yen Chang, Ya-Chun Yang, and Pei-Kwei Tsay. "Effects of Cryopreservation Duration on the Outcome of Single-Unit Cord Blood Transplantation." Cell Transplantation 27, no. 3 (March 2018): 515–19. http://dx.doi.org/10.1177/0963689717753187.

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Cryopreservation is widely used in umbilical cord blood (UCB) banking, yet its impact on progenitor cell function remains largely unaddressed. It is unknown whether long-term cryopreservation affects UCB transplantation outcomes. Herein, we evaluated the impact of UCB age on clinical outcomes and investigated the effect of cryopreservation duration of UCB on hematopoietic potency in 91 patients receiving single cord blood transplantations. UCB cryopreservation duration was 0.7 to 13.4 y. The most common indication of transplant was thalassemia (48%). There was no significant association between cryopreservation duration and neutrophil engraftment probability ( P = 0.475). Cryopreservation duration did not affect the post-thaw viability and subsequent neutrophil engraftment rate. Therefore, UCB units can undergo cryopreservation for at least 8 y with no impact on clinical outcomes.

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Pham, Phuc Van, Tam Thanh Nguyen, Nhung Thi Hong Vuong, Tuyet Thi Bach Duong, and Ngoc Kim Phan. "INVESTIGATING COOLING RATE AND FETAL BOVINE SERUM CONCENTRATION ON SURVIVAL AND STEMNESS OF MESENCHYMAL STEM CELLS AFTER CRYOPRESERVATION." Science and Technology Development Journal 12, no. 9 (May 15, 2009): 12–22. http://dx.doi.org/10.32508/stdj.v12i9.2281.

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Mesenchymal stem cells (MSCs) can be derived from many different sources. Umbilical cord blood is a rich source of MSCs. The cryopreservation of MSCs that MSCs are still alive and differentiate into many different kinds of functional cells is very important. The aims of this research are to identify ratio of alive and dead cells as well as stemness of them after thaw. The results showed that the stemness was not affected by cryopreservative protocols or media. All cells being alive after thaw could form colonies and differentiate into adipocytes and osteoblasts. Ratio of alive and dead cells was affected very much by cryopreservative protocols and media.

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Yang, Jing, Lei Gao, Min Liu, Xiaojie Sui, Yingnan Zhu, Chiyu Wen, and Lei Zhang. "Advanced Biotechnology for Cell Cryopreservation." Transactions of Tianjin University 26, no. 6 (December 28, 2019): 409–23. http://dx.doi.org/10.1007/s12209-019-00227-6.

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AbstractCell cryopreservation has evolved as an important technology required for supporting various cell-based applications, such as stem cell therapy, tissue engineering, and assisted reproduction. Recent times have witnessed an increase in the clinical demand of these applications, requiring urgent improvements in cell cryopreservation. However, cryopreservation technology suffers from the issues of low cryopreservation efficiency and cryoprotectant (CPA) toxicity. Application of advanced biotechnology tools can significantly improve post-thaw cell survival and reduce or even eliminate the use of organic solvent CPAs, thus promoting the development of cryopreservation. Herein, based on the different cryopreservation mechanisms available, we provide an overview of the applications and achievements of various biotechnology tools used in cell cryopreservation, including trehalose delivery, hydrogel-based cell encapsulation technique, droplet-based cell printing, and nanowarming, and also discuss the associated challenges and perspectives for future development.

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Haider, Patrick, Timothy Hoberstorfer, Manuel Salzmann, Michael B. Fischer, Walter S. Speidl, Johann Wojta, and Philipp J. Hohensinner. "Quantitative and Functional Assessment of the Influence of Routinely Used Cryopreservation Media on Mononuclear Leukocytes for Medical Research." International Journal of Molecular Sciences 23, no. 3 (February 7, 2022): 1881. http://dx.doi.org/10.3390/ijms23031881.

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Quantitative and functional analysis of mononuclear leukocyte populations is an invaluable tool to understand the role of the immune system in the pathogenesis of a disease. Cryopreservation of mononuclear cells (MNCs) is routinely used to guarantee similar experimental conditions. Immune cells react differently to cryopreservation, and populations and functions of immune cells change during the process of freeze–thawing. To allow for a setup that preserves cell number and function optimally, we tested four different cryopreservation media. MNCs from 15 human individuals were analyzed. Before freezing and after thawing, the distribution of leukocytes was quantified by flow cytometry. Cultured cells were stimulated using lipopolysaccharide, and their immune response was quantified by flow cytometry, quantitative polymerase chain reaction (qPCR), and enzyme-linked immunosorbent assay (ELISA). Ultimately, the performance of the cryopreservation media was ranked. Cell recovery and viability were different between the media. Cryopreservation led to changes in the relative number of monocytes, T cells, B cells, and their subsets. The inflammatory response of MNCs was altered by cryopreservation, enhancing the basal production of inflammatory cytokines. Different cryopreservation media induce biases, which needs to be considered when designing a study relying on cryopreservation. Here, we provide an overview of four different cryopreservation media for choosing the optimal medium for a specific task.

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Pegg, David E., Monica C. Wusteman, and Lihong Wang. "Cryopreservation of articular cartilage. Part 1: Conventional cryopreservation methods." Cryobiology 52, no. 3 (June 2006): 335–46. http://dx.doi.org/10.1016/j.cryobiol.2006.01.005.

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Arnold, Stacy, and David Gale. "26. Optimizing tissue cryopreservation techniques for cryopreservation solution modifications." Cryobiology 66, no. 3 (June 2013): 349. http://dx.doi.org/10.1016/j.cryobiol.2013.02.032.

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Alnemari, Roaa, Pavithra Sukumar, Muhammedin Deliorman, and Mohammad A. Qasaimeh. "Cryopreservation: Paper‐Based Cell Cryopreservation (Adv. Biosys. 3/2020)." Advanced Biosystems 4, no. 3 (March 2020): 2070033. http://dx.doi.org/10.1002/adbi.202070033.

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Tucker, Michael, Paula Morton, and Juergen Liebermann. "Human oocyte cryopreservation: a valid alternative to embryo cryopreservation?" European Journal of Obstetrics & Gynecology and Reproductive Biology 113 (April 2004): S24—S27. http://dx.doi.org/10.1016/j.ejogrb.2003.11.006.

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Vanderzwalmen, Pierre, Fabien Ectors, Yannis Panagiotidis, Maximilian Schuff, Maximilian Murtinger, and Barbara Wirleitner. "The Evolution of the Cryopreservation Techniques in Reproductive Medicine—Exploring the Character of the Vitrified State Intra- and Extracellularly to Better Understand Cell Survival after Cryopreservation." Reproductive Medicine 1, no. 2 (September 17, 2020): 142–57. http://dx.doi.org/10.3390/reprodmed1020011.

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Nowadays, cryopreservation of gametes and embryos is a fundamental, integral, and indispensable part of infertility treatment or fertility preservation. Cryopreservation is not only needed for the policy of single embryo transfer and cryopreservation of surplus embryos, but for deferring embryo transfer in the case of ovarian hyperstimulation syndrome, uterine pathologies, and suboptimal endometrium built-up or when preimplantation genetic testing is needed. Several current strategies in assisted reproduction technology (ART) would be inconceivable without highly efficient cryopreservation protocols. Nevertheless, cryopreservation hampered for a long time, especially in terms of low survival rates after freezing and thawing. Only the technical progress during the last decades, namely, in regard to the implementation and advancement of vitrification, leveraged its application, and thus, even allows the cryopreservation of human oocytes—a process that is far from being easy. This review aims to provide a deeper insight into the physical processes of cryopreservation and to explore the character of the vitrified state in the extra and intracellular milieu in order to demonstrate that the common denominator to all cryopreservation procedures is the establishment of an intracellular amorphous condition that hinders the likelihood of crystallization.

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Moraes, Rodrigo Miranda, Fernanda Carlota Nery, Mayara Caroline Carvalho Pinto, Renato Paiva, and Sandro Barbosa. "Conservation of Hibiscus acetosella germplasm by seed cryopreservation." 2019 13, (03) 2019 (May 20, 2019): 372–79. http://dx.doi.org/10.21475/ajcs.19.13.03.p1209.

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Hibiscus acetosella (Malvaceae) is a shrub of great importance for landscaping, food and medicinal purposes. The objective of this study was to preserve H. acetosella germplasm by seed cryopreservation. Half of the seed batch was scarified and the other half was kept intact. Cryopreservation occurred by immersion in liquid nitrogen for 1 hour. Moisture content (MC%), germination percentage (G%), germination speed index (GSI), normal seedling formation (NS%), shoot length (SL), dry matter (DM), biometry and plant survival were evaluated after treatment. MC% ranged between 7.7% and 6.65% in intact and scarified seeds, respectively. Scarification raised G% and GSI compared to intact seeds. Intact and scarified seeds had 100% and 70% NS%, respectively, when not cryopreserved. Cryopreservation reduced NS% to 62% and 12.75%, respectively. The highest SL was observed in intact and non-cryopreserved seeds, with an average of 10.21 cm in height. However, the cryopreservation of intact seeds reduced SL by about 50%, and scarification led to a further reduction, either with (3.32 cm) or without (2.47 cm) cryopreservation. Seedlings from intact and non-cryopreserved seeds showed higher DM in relation to seedlings from cryopreserved seeds. The association of cryopreservation and scarification further reduced DM. The cryopreservation of intact seeds yielded 100% survival at the end of the acclimatization process. However, cryopreservation of scarified seeds reduced the survival percentage to 15%. Changes in color were observed for seeds scarified and subjected to cryopreservation. Thus, cryopreservation is considered an efficient technique for the conservation of intact H. acetosella seeds in the long term.

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Sisková, Ingrid, Lenka Mekiňová, Robert Hudeček, Michal Ješeta, and Soňa Šimová. "Methods and techniques of fertility preservation in patients with endometriosis." Česká gynekologie 88, no. 6 (December 20, 2023): 454–58. http://dx.doi.org/10.48095/cccg2023454.

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Objective: Endometriosis is a chronic disease with a relatively high prevalence in the female population. Both the disease itself and its surgical treatment can adversely affect the fertility of patients. For this reason, endometriosis is offered as a possible indication for fertility preservation by cryopreservation methods. The aim of this paper is to present the current knowledge on the options of fertility preservation in this subpopulation. Methods: Search of relevant literature in PubMed/Medline, Web of Science and Scopus databases. Results: Fertility preservation by cryopreservation methods has so far been used mainly in the care of women with cancer. With increasing experience, the effectiveness and availability of these methods have increased significantly and the indication spectrum has been extended to selected benign diseases such as endometriosis. Three techniques are currently established in practice: embryo cryopreservation, oocyte cryopreservation and ovarian tissue cryopreservation. Oocyte cryopreservation is the most commonly used technique, since it is the most advantageous for the patient and, according to the available data, is an effective way to increase the chances of future pregnancy for patients with endometriosis The purpose is to realize the protection of reproduction before the planned operation. Conclusion: The diagnosis of endometriosis negatively affects the fertility of women. For some patients, the solution is fertility preservation by cryopreservation methods. Further clinical studies are needed to define exact, practically applicable indication criteria, potential risks of procedures and their benefits and cost-effectiveness. Key words: fertility preservation – endometriosis – oocyte cryopreservation – embryo cryopreservation – ovarian tissue cryopreservation

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Karabulut, Seda, Asuman Demiroğlu-Zergeroğlu, Elif Yılmaz, Pelin Kutlu, and İlknur Keskin. "Effects of human sperm cryopreservation on apoptotic markers in normozoospermic and non-normozoospermic patients." Zygote 26, no. 4 (August 2018): 308–13. http://dx.doi.org/10.1017/s0967199418000254.

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SummaryThe negative effects of cryopreservation on sperm parameters are well documented but little information is known about molecular basis of the process. The aim of the present study was to investigate the possible effects of sperm cryopreservation on main apoptotic signs including DNA fragmentation and caspase-3 activation and to determine if these effects vary according to sperm parameters. Sperm samples of 72 patients were cryopreserved. The patients were sub-grouped as normozoospermic or non-normozoospermic patients according to their semen parameters. DNA fragmentation rates and caspase-3 activation levels were analyzed before and after cryopreservation in both groups. Mean DNA fragmentation rate was increased significantly from 23.98% in neat semen samples to 27.34% after cryopreservation (P = 0.03). DNA fragmentation rates were slightly higher in non-normozoospermic patients compared with the normozoospermic patients in both the neat semen and after cryopreservation (23.25 and 24.71% vs. 26.32 and 28.36%, respectively) although the difference obtained were not statistically significant. An increasing trend for caspase-3 activations (0.093 vs. 0.116) was observed after cryopreservation but the differences were not statistically significant. Caspase-3 activation was found to be slightly higher in non-normozoospermic patients both in the neat semen and after cryopreservation compared with the normozoospermic patients but the differences were not statistically significant. Caspase-3 expression was also shown using immunocytochemistry in both fresh ejaculated sperm and thawed sperm after cryopreservation but at different localizations. The cryopreservation process had detrimental effects on sperm quality but the quality of the sperm samples was not adversely effective for the apoptotic markers including DNA fragmentation and caspase-3 activation patterns. In fact, it was the cryopreservation process itself that adversely effected the above apoptotic markers and apoptosis. It was concluded therefore that sperm cell cryopreservation triggers apoptosis after thawing and this process adversely affects semen parameters.

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Ibrahim, Mohammad A. "Bull sperm cryopreservation: An overview on the current status and future perspectives." German Journal of Veterinary Research 4, no. 1 (January 2024): 9–22. http://dx.doi.org/10.51585/gjvr.2024.1.0071.

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Cryopreservation refers to freezing cells or tissues at extremely low temperatures, allowing them to be stored for extended periods while maintaining viability. Cryopreserved bull semen has become an essential tool in cattle breeding programs and commercial cattle production systems. This review provides a detailed analysis of the current methods and challenges in preserving bull sperm using cryopreservation. We explore the effects of cryopreservation on sperm cells, the role of different cryoprotectants, as well as the progress made in the analysis of bull semen. It also highlights the impact of the freezing process on sperm morphology and functionality, emphasizing the importance of optimizing cryopreservation techniques to maintain sperm fertility and viability. The article underscores the significance of cryopreservation technology in cattle genetics and breeding and suggests future research to enhance cryopreservation techniques.

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Grandhaye, Jérémy, Agnieszka Partyka, Zuzanna Ligocka, Agata Dudek, Wojciech Niżański, Eric Jeanpierre, Anthony Estienne, and Pascal Froment. "Metformin Improves Quality of Post-Thaw Canine Semen." Animals 10, no. 2 (February 12, 2020): 287. http://dx.doi.org/10.3390/ani10020287.

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Sperm cryopreservation is an assisted reproductive technique routinely used in canine species for genetic conservation. However, during cryopreservation, the DNA damages are still elevated, limiting the fertilization rate. The present study was conducted to evaluate whether supplementation of canine semen extender with a molecule limiting the metabolic activities can improve the quality of frozen-thawed canine spermatozoa. We used metformin, known to limit the mitochondrial respiratory and limit the oxidative stress. Before and during the freezing procedure, metformin (50µM and 500µM) has been added to the extender. After thawing, sperm exposed to metformin conserved the same viability without alteration in the membrane integrity or acrosome reaction. Interestingly, 50µM metformin improved the sperm motility in comparison to the control, subsequently increasing mitochondrial activity and NAD+ content. In addition, the oxidative stress level was reduced in sperm treated with metformin improving the sperm quality as measured by a different molecular marker. In conclusion, we have shown that metformin is able to improve the quality of frozen-thawed dog semen when it is used during the cryopreservative procedure.

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Khodko, Oleksiy. "Role of Liquid-Liquid Phase Transitions in Mechanism of Erythrocyte Protection During Cooling with CRIHBT-115 Cryopreservative Agent." Problems of Cryobiology and Cryomedicine 31, no. 3 (September 25, 2021): 236–48. http://dx.doi.org/10.15407/cryo31.03.236.

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The presence in the system of critical liquid-liquid phase transition (PT) by the mechanism, resulting in formation of dispersion system, namely high-concentrated emulsion, has been established here during cooling when using polarized light microscopy and fixation of critical opalescence phenomenon in erythrocyte concentrate with glycerol-containing cryopreservative agent, designed at the Central Research Institute of Haematology and Blood Transfusion (Russia) (CRIHBT-115 ). The studied cryobiological system displayed no signs of crystallization. A phase behaviour of cryopreservative and supernatant has been studied during cooling-warming cycle. Changes in the volume of cryopreservative and erythrocyte concentrate were comparatively and qualitatively evaluated during cooling. The mechanism of protective action of cryopreservation solution has been determined. The similarity between physical and chemical processes during cooling-warming of erythrocyte cytoplasm and garlic meristem cells (germinal plant tissue) when entering cold anabiosis has been established.

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Matsushita, Takakazu, Toshikazu Yagi, Joseph A. Hardin, Jennifer D. Cragun, Frank W. Crow, H. Robert Bergen, Gregory J. Gores, and Scott L. Nyberg. "Apoptotic Cell Death and Function of Cryopreserved Porcine Hepatocytes in a Bioartificial Liver." Cell Transplantation 12, no. 2 (March 2003): 109–21. http://dx.doi.org/10.3727/000000003108746696.

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We have previously shown that cryopreservation leads to increased apoptotic death of porcine hepatocytes intended for use in a bioartificial liver (BAL). This study was designed to determine if a broad-spectrum caspase inhibitor, IDN–1965, reduced apoptosis and increased function of cryopreserved porcine hepatocytes in static culture or in a BAL. Porcine hepatocytes were studied immediately after isolation and after 2 weeks of cryopreservation in liquid nitrogen using medium supplemented with 25 μmol/L IDN-1965 or vehicle. Both apoptotic and necrotic cells were observed in cultures of fresh and cryopreserved hepatocytes, but the percentage of apoptotic cells increased after cryopreservation. Cryopreservation in IDN-1965 improved hepatocyte viability and reduced apoptotic cell death determined by TUNEL assay. Cryopreservation of hepatocytes in IDN-1965 was also associated with reduced caspase 3-like activity, decreased release of cytochrome c from mitochondria, and a slower decline in mitochondrial membrane potential after thawing. These markers of apoptosis were lowest after cryopreservation when IDN-1965 was added to both the culture and cryopreservation medium. Functional markers of hepatocyte activity (albumin production, diazepam metabolism, urea production) were also increased after cryopreservation and culture of hepatocytes in medium supplemented with 25 μmol/L IDN-1965. Cryopreservation of porcine hepatocytes in the presence of caspase inhibitor IDN-1965 was associated with reduced apoptosis and improved function of porcine hepatocytes in both static culture and a perfused BAL. These data demonstrate that inhibition of apoptosis also preserves cell function.

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Xu, Feng, Sangjun Moon, Xiaohui Zhang, Lei Shao, Young Seok Song, and Utkan Demirci. "Multi-scale heat and mass transfer modelling of cell and tissue cryopreservation." Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences 368, no. 1912 (February 13, 2010): 561–83. http://dx.doi.org/10.1098/rsta.2009.0248.

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Cells and tissues undergo complex physical processes during cryopreservation. Understanding the underlying physical phenomena is critical to improve current cryopreservation methods and to develop new techniques. Here, we describe multi-scale approaches for modelling cell and tissue cryopreservation including heat transfer at macroscale level, crystallization, cell volume change and mass transport across cell membranes at microscale level. These multi-scale approaches allow us to study cell and tissue cryopreservation.

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NUKARI, A., M. UOSUKAINEN, and V.-M. ROKKA. "Cryopreservation techniques and their application in vegetatively propagated crop plants in Finland." Agricultural and Food Science 18, no. 2 (December 4, 2008): 117. http://dx.doi.org/10.2137/145960609789267506.

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Cryopreservation protocols have been introduced as techniques for germplasm preservation of vegetatively propagated horticultural and staple food crops. In Finland, cryopreservation has been studied since 1990s, beginning with cryopreservation of forest tree breeding material and since 2004 on cryopreservation of genetic resources of horticultural plants and potato. Priority was given to cryopreservation of raspberry (Rubus ideaus L.), strawberry (Fragaria x ananassa Duch.) and potato (Solanum tuberosum L.) and the possibility to use cryotherapy in eradication of raspberry bushy dwarf virus (RBDV) from in vitro cultures were studied on raspberry. Modified droplet vitrification cryopreservation protocols were designed for raspberry and strawberry and cryotherapy combined with thermotherapy was proven to be a successful application to eliminate RBDV from infected raspberries. Cryotherapy method can be applied for a large scale elimination of viruses from plant germplasm and from candidate nuclear stock in a certified plant production scheme. Routine use of cryotechniques in germplasm preservation of vegetatively propagated horticultural plants was started. Besides for long term germplasm preservation, cryopreservation techniques can be applied also for maintenance of mother stocks in certified plant production schemes and in commercial plant production. Cryopreservation of potato shoot tips needs additional detailed research to obtain sufficient recovery and regrowth rates.;

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Ribeiro, João C., David F. Carrageta, Raquel L. Bernardino, Marco G. Alves, and Pedro F. Oliveira. "Aquaporins and Animal Gamete Cryopreservation: Advances and Future Challenges." Animals 12, no. 3 (February 2, 2022): 359. http://dx.doi.org/10.3390/ani12030359.

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Cryopreservation is globally used as a method for long-term preservation, although freeze-thawing procedures may strongly impair the gamete function. The correct cryopreservation procedure is characterized by the balance between freezing rate and cryoprotective agents (CPAs), which minimizes cellular dehydration and intracellular ice formation. For this purpose, osmoregulation is a central process in cryopreservation. During cryopreservation, water and small solutes, including penetrating cryoprotective agents, cross the plasma membrane. Aquaporins (AQPs) constitute a family of channel proteins responsible for the transport of water, small solutes, and certain gases across biological membranes. Thirteen homologs of AQPs (AQP0-12) have been described. AQPs are widely distributed throughout the male and female reproductive systems, including the sperm and oocyte membrane. The composition of the male and female gamete membrane is of special interest for assisted reproductive techniques (ART), including cryopreservation. In this review, we detail the mechanisms involved in gamete cryopreservation, including the most used techniques and CPAs. In addition, the expression and function of AQPs in the male and female gametes are explored, highlighting the potential protective role of AQPs against damage induced during cryopreservation.

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Amini, Mohammad, and James D. Benson. "Technologies for Vitrification Based Cryopreservation." Bioengineering 10, no. 5 (April 23, 2023): 508. http://dx.doi.org/10.3390/bioengineering10050508.

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Cryopreservation is a unique and practical method to facilitate extended access to biological materials. Because of this, cryopreservation of cells, tissues, and organs is essential to modern medical science, including cancer cell therapy, tissue engineering, transplantation, reproductive technologies, and bio-banking. Among diverse cryopreservation methods, significant focus has been placed on vitrification due to low cost and reduced protocol time. However, several factors, including the intracellular ice formation that is suppressed in the conventional cryopreservation method, restrict the achievement of this method. To enhance the viability and functionality of biological samples after storage, a large number of cryoprotocols and cryodevices have been developed and studied. Recently, new technologies have been investigated by considering the physical and thermodynamic aspects of cryopreservation in heat and mass transfer. In this review, we first present an overview of the physiochemical aspects of freezing in cryopreservation. Secondly, we present and catalog classical and novel approaches that seek to capitalize on these physicochemical effects. We conclude with the perspective that interdisciplinary studies provide pieces of the cryopreservation puzzle to achieve sustainability in the biospecimen supply chain.

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Ranjan, R., M. Kumar, C. Gangwar, and S. D. Kharche. "Developments in Goat Semen Cryopreservation." Animal Reproduction Update 1, no. 1 (2021): 41–45. http://dx.doi.org/10.48165/aru.2021.1205.

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Sperm cryopreservation simplifies its storage for longer time for use in artificial insemination and assisted reproductive technologies. This technique is also important for breed conservation process and has paved the way for other reproductive biotechnologies. Despite the significant progress in the field, frozen-thawed sperm have enormous inconsistency in fertility rates. It is well known fact that semen cryopreservation exhibited detrimental effects on post-thaw semen motility, plasma membrane, acrosomal status and DNA integrity which ultimately effect the fertility outcome. In addition, several attributes are responsible for low quality of goat cryopreserved semen such as breeds, seasons, management practices and cryopreservation protocols. The aim of the review article is to give an insight of distant features of goat semen cryopreservation as well as recent development in goat semen cryopreservation. It also provides concise information on progress made in the advancement in the semen extender development and cryopreservation of goat sperm.

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Zong, Yunhe, Yunlei Li, Yanyan Sun, Gamal M. K. Mehaisen, Tianxiao Ma, and Jilan Chen. "Chicken Sperm Cryopreservation: Review of Techniques, Freezing Damage, and Freezability Mechanisms." Agriculture 13, no. 2 (February 14, 2023): 445. http://dx.doi.org/10.3390/agriculture13020445.

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Ex situ preservation is an important method in the preservation of chickens, and cryopreservation of semen is the only method for gamete preservation at present. During the last two decades, many studies have been performed to develop standard chicken semen cryopreservation technology and achieve great progress. Many attempts and methods were investigated to adapt subspecies or different breeds. In this paper, we firstly reviewed the main factors affecting cryopreservation of chicken sperm, including the unique structure and characteristics of the spermatozoa. Secondly, the studies on key points of the chicken sperm cryopreservation technology, including semen dilution, cryoprotectants, equilibration time, packaging types, and freezing and thawing rates were summarized to generate the optimal parameters. Then, the mechanism underlying freezing damage and freezability revealed by recent omics methods relevant to the efficiency of cryopreservation were discussed. This review will provide relevant reference for the future investigation of poultry semen cryopreservation technology.

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McLemore, T. L., and T. J. Kuehl. "Measurement of Viability and Aryl Hydrocarbon Hydroxylase Activity in Cryopreserved Baboon Peripheral Blood Lymphocytes and Pulmonary Alveolar Macrophages." Journal of the American College of Toxicology 4, no. 2 (March 1985): 193–99. http://dx.doi.org/10.3109/10915818509014514.

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Pulmonary alveolar macrophages (PAMs) and peripheral blood lymphocytes obtained from healthy nonsmoking baboons were evaluated for the ability to retain viability and aryl hydrocarbon hydroxylase (AHH) activity following cryopreservation for 2, 4, 6, 8, and 12 week intervals. Viability was measured (by trypan blue dye exclusion) in lymphocytes following 2, 4, 6, 8, and 12 weeks of cryopreservation. No significant decrease in viability was observed (P > 0.05 for all comparisons; nonpaired 2-tailed, t-test; n = 22), with lymphocytes retaining 88% of their original viability following 12 weeks of cryopreservation. Similarly, a slight, but insignificant decrease in PAM viability was observed following cryopreservation for 2, 4, 6, and 8 week intervals (P > 0.05 in all instances), with PAMs retaining 93.5% of the original 0 time viability. AHH activity was also evaluated in PAMs and lymphocytes following cryopreservation. Lymphocyte noninduced and BA-induced AHH activities were slightly but not significantly decreased after 2, 4, 6, 8, and 12 week cryopreservation intervals (P > 0.05 for all comparisons). Similarly, AHH activity in cultured PAMs was not significantly decreased following cryopreservation for 2, 4, 6, and 8 week intervals (P > 0.05 for all noninduced and BA-induced AHH levels assayed at the various time intervals; n = 8). Even following cryopreservation periods, for 8 or 12 week intervals, PAMs and lymphocytes retained 91% and 90%, respectively, of the original BA-induced AHH activity. These data demonstrate that baboon PAMs and lymphocytes are capable of undergoing cryopreservation while maintaining viability and AHH activity.

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Lu, Jingjing, Xuezi Tian, and Zhaochen Wang. "Latent class analysis of Chinese healthcare providers’ attitudes towards oocyte cryopreservation: a cross-sectional study." BMJ Open 14, no. 3 (March 2024): e076680. http://dx.doi.org/10.1136/bmjopen-2023-076680.

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ObjectivesThe present study was designed to examine the attitudes towards oocyte cryopreservation among healthcare providers working in hospitals across specialties and potential influencing factors.DesignA cross-sectional study.SettingThe questionnaire was distributed among Chinese healthcare providers via the Credamo platform.ParticipantsThere were 877 respondents recruited from 8 April to 8 May 2022, among whom 160 were identified as unqualified because of inconsistency between the IP and work addresses.Outcome measuresIndividual attitudes towards oocyte cryopreservation under four different settings, familiarity with oocyte cryopreservation and perceived risks about oocyte cryopreservation of healthcare providers were measured using a self-designed questionnaire.ResultsThere were 877 respondents recruited, and 717 were identified as qualified respondents. Two latent classes of healthcare providers characterised by different attitudes towards oocyte cryopreservation under four different settings were identified, the supportive and reluctant. Familiarity with oocyte cryopreservation had a significant direct effect on perceived risks, with better familiarity predicting lower perceived risks (β=−0.102, p<0.05). Perceived risks showed a significant direct effect on participants’ attitudes towards oocyte cryopreservation, with higher perceived risks predicting a more reluctant attitude (β=0.165, p<0.001).ConclusionsThe majority of healthcare providers held a reluctant attitude towards oocyte cryopreservation of unmarried women for non-medical reasons, which might relate to their worries about the risks to offspring’s health and lack of knowledge about a reproductive technique.

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Afriani, Dian, Kartini Eriani, Zainal Abidin Muchlisin, and Iwan Hasri. "A short review of discovery and development of fish sperm cryopreservation." Depik 10, no. 1 (February 12, 2021): 11–16. http://dx.doi.org/10.13170/depik.10.1.18794.

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Global biodiversity, especially fish, has experienced a decline, this occurs as a result of over-exploitation, the presence of introduced fish species and climate change. This condition makes researchers look for solutions to overcome these problems by using cryopreservation techniques. The main purpose of cryopreservation is to store, maintain, and ensure the survival of genetic material, so that using cryopreservation techniques can maintain the viability and function of gamete cells both immunologically, biologically and physiologically. The success of the cryopreservation technique has made this technique widely developed in various species of living organism including fish. This article summarizes and reviews the history of the development of cryopreservation of animal species with specific focus on fish.Keywords:CryopreservationHistoryDepikEndemic species

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Bradaï, Fatiha, and Carolina Sánchez-Romero. "Effect of Cryopreservation on the Ex Vitro Establishment of Olive Plants Regenerated via Somatic Embryogenesis." Plants 10, no. 2 (February 19, 2021): 396. http://dx.doi.org/10.3390/plants10020396.

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Cryopreservation is considered the best technique for the safe, long-term conservation of embryogenic cultures. However, before integrating it into a somatic embryogenesis system, the influence of cryopreservation on the final production of plants should be investigated. The objective of this investigation was to evaluate the effect of cryopreservation on the regeneration performance of olive embryogenic cultures as well as on the quality of the plants obtained and their response to ex vitro establishment. In order to analyze the influence of the genotype, all the investigations were carried out in two genetically distinct embryogenic lines. The results obtained revealed no variation in the regeneration potential or the quality of the regenerated plants due to cryopreservation. The subsequent multiplication, rooting, and acclimatization steps were not influenced by cryopreservation either, although a significant genotype × cryopreservation interaction was found for shoot length during the multiplication step. The genotype played an important role, determining the quality of the regenerated plants and some aspects of the multiplication and rooting phases. This investigation revealed that the droplet-vitrification procedure optimized for the cryopreservation of olive somatic embryos can be efficiently used for the long-term conservation of olive embryogenic lines.

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Saha, Amit, Mohammad Asaduzzaman, and Farida Yeasmin Bari. "Cryopreservation Techniques for Ram Sperm." Veterinary Medicine International 2022 (April 30, 2022): 1–16. http://dx.doi.org/10.1155/2022/7378379.

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Germplasm storage and transportation in artificial insemination (AI) and other advanced technologies are facilitated by cryopreservation. In reproduction, the cryopreservation of sperm allows it to be transported across vast distances and used even after the sire’s death. However, the technique of cryopreservation might damage sperm and limit their activity. Several cryobiological investigations have reported that the integrity of the sperm membrane is frequently involved in the physical and biological elements that affect sperm survival at low temperatures during the cryopreservation process. However, successful cryopreservation of ram sperm is still a work in progress because a considerable percentage of sperm do not survive the freezing and thawing process. Sperms are destroyed during cryopreservation of semen due to varying concentrations of cryoprotective chemicals and if semen is not cooled at optimal cooling rates. Hence, it is crucial to know the optimum cooling rates with freezing and thawing protocols for maximum recovery of viable and functional sperm cells for a successful cryo-freezing of ram spermatozoa. Therefore, the current study compiled and compared the research on the impact of different cryopreservation procedures, cooling rates, equilibration time, and thawing protocols on post-thaw ram semen quality.

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Journal articles on the topic 'Cryopreservation'

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