Статті в журналах: "Tissue culture"

1

Jia, Zhidong, Yuan Cheng, Xinan Jiang, Chengyan Zhang, Gaoshang Wang, Jiecheng Xu, Yang Li, Qing Peng, and Yi Gao. "3D Culture System for Liver Tissue Mimicking Hepatic Plates for Improvement of Human Hepatocyte (C3A) Function and Polarity." BioMed Research International 2020 (March 4, 2020): 1–22. http://dx.doi.org/10.1155/2020/6354183.

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In vitro 3D hepatocyte culture constitutes a core aspect of liver tissue engineering. However, conventional 3D cultures are unable to maintain hepatocyte polarity, functional phenotype, or viability. Here, we employed microfluidic chip technology combined with natural alginate hydrogels to construct 3D liver tissues mimicking hepatic plates. We comprehensively evaluated cultured hepatocyte viability, function, and polarity. Transcriptome sequencing was used to analyze changes in hepatocyte polarity pathways. The data indicate that, as culture duration increases, the viability, function, polarity, mRNA expression, and ultrastructure of the hepatic plate mimetic 3D hepatocytes are enhanced. Furthermore, hepatic plate mimetic 3D cultures can promote changes in the bile secretion pathway via effector mechanisms associated with nuclear receptors, bile uptake, and efflux transporters. This study provides a scientific basis and strong evidence for the physiological structures of bionic livers prepared using 3D cultures. The systems and cultured liver tissues described here may serve as a better in vitro 3D culture platform and basic unit for varied applications, including drug development, hepatocyte polarity research, bioartificial liver bioreactor design, and tissue and organ construction for liver tissue engineering or cholestatic liver injury.

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Shrestha, Sunil, Vinod Kumar Reddy Lekkala, Prabha Acharya, Darshita Siddhpura, and Moo-Yeal Lee. "Recent advances in microarray 3D bioprinting for high-throughput spheroid and tissue culture and analysis." Essays in Biochemistry 65, no. 3 (August 2021): 481–89. http://dx.doi.org/10.1042/ebc20200150.

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Abstract Three-dimensional (3D) cell culture in vitro has proven to be more physiologically relevant than two-dimensional (2D) culture of cell monolayers, thus more predictive in assessing efficacy and toxicity of compounds. There have been several 3D cell culture techniques developed, which include spheroid and multicellular tissue cultures. Cell spheroids have been generated from single or multiple cell types cultured in ultralow attachment (ULA) well plates and hanging droplet plates. In general, cell spheroids are formed in a relatively short period of culture, in the absence of extracellular matrices (ECMs), via gravity-driven self-aggregation, thus having limited ability to self-organization in layered structure. On the other hand, multicellular tissue cultures including miniature tissues derived from pluripotent stem cells and adult stem cells (a.k.a. ‘organoids’) and 3D bioprinted tissue constructs require biomimetic hydrogels or ECMs and show highly ordered structure due to spontaneous self-organization of cells during differentiation and maturation processes. In this short review article, we summarize traditional methods of spheroid and multicellular tissue cultures as well as their technical challenges, and introduce how droplet-based, miniature 3D bioprinting (‘microarray 3D bioprinting’) can be used to improve assay throughput and reproducibility for high-throughput, predictive screening of compounds. Several platforms including a micropillar chip and a 384-pillar plate developed to facilitate miniature spheroid and tissue cultures via microarray 3D bioprinting are introduced. We excluded microphysiological systems (MPSs) in this article although they are important tissue models to simulate multiorgan interactions.

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Peel, Trisha N., Tim Spelman, Brenda L. Dylla, John G. Hughes, Kerryl E. Greenwood-Quaintance, Allen C. Cheng, Jayawant N. Mandrekar, and Robin Patel. "Optimal Periprosthetic Tissue Specimen Number for Diagnosis of Prosthetic Joint Infection." Journal of Clinical Microbiology 55, no. 1 (November 2, 2016): 234–43. http://dx.doi.org/10.1128/jcm.01914-16.

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ABSTRACTWe recently demonstrated improved sensitivity of prosthetic joint infection (PJI) diagnosis using an automated blood culture bottle system for periprosthetic tissue culture [T. N. Peel et al., mBio 7(1):e01776-15, 2016,https://doi.org/10.1128/mBio.01776-15]. This study builds on the prior research by examining the optimal number of periprosthetic tissue specimens required for accurate PJI diagnosis. Current guidelines recommend five to six, which is impractical. We applied Bayesian latent class modeling techniques for estimating diagnostic test properties of conventional culture techniques (aerobic and anaerobic agars and thioglycolate broth) compared to inoculation into blood culture bottles. Conventional, frequentist receiver operating characteristic curve analysis was conducted as a sensitivity analysis. The study was conducted at Mayo Clinic, Rochester, MN, from August 2013 through April 2014 and included 499 consecutive patients undergoing revision arthroplasty from whom 1,437 periprosthetic tissue samples were collected and processed. For conventional periprosthetic tissue culture techniques, the greatest accuracy was observed when four specimens were obtained (91%; 95% credible interval, 77 to 100%), whereas when using inoculation of periprosthetic tissues into blood culture bottles, the greatest accuracy of diagnosis was observed when three specimens were cultured (92%; 95% credible intervals, 79 to 100%). Results of this study show that the greatest accuracy of PJI diagnosis is obtained when three periprosthetic tissue specimens are obtained and inoculated into blood culture bottles or four periprosthetic tissue specimens are obtained and cultured using standard plate and broth cultures. Increasing the number of specimens to five or more, per current recommendations, does not improve accuracy of PJI diagnosis.

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Şükrüoğlu Erdoğan, Özge, Seda Kılıç Erciyas, Ayhan Bilir, Şeref Buğra Tunçer, Demet Akdeniz Ödemiş, Sıdıka Kurul, Hasan Karanlık, Neslihan Cabıoğlu, and Hülya Yazıcı. "Methylation Changes of Primary Tumors, Monolayer, and Spheroid Tissue Culture Environments in Malignant Melanoma and Breast Carcinoma." BioMed Research International 2019 (January 17, 2019): 1–9. http://dx.doi.org/10.1155/2019/1407167.

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Epigenetic changes have major role in the normal development and programming of gene expression. Aberrant methylation results in carcinogenesis. The primary objective of our study is to determine whether primary tumor tissue and cultured tumor cells in 2D and 3D tissue culture systems have the same methylation signature forPAX5,TMPRSS2, andSBDS. These findings will play an important role in developing in vitro model system to understand the effect of methylation inhibitors on primary tumor tissue. In a previous studyPAX5,TMPRSS2, andSBDSgenes that we are investigating were reported to be methylated more than 60% in breast cancer and malignant melanoma cell lines. However, these genes have never been studied in primary tumor tissues. Thus, primary tumor tissues of breast cancer and malignant melanoma were first grown in 2D and 3D cultures. Then these two types of tumor tissues and their 2D and 3D cultures were investigated for changes considering methylation levels inPAX5,TMPRSS2, andSBDSgenes using real-time polymerase chain reaction. No differences were observed in the primary tissues and culture systems for bothPAX5andTMPRSS2in malignant melanoma tissues. We found thatPAX5gene was an efficient marker to measure the effects of methylation inhibitors for in vitro systems for malignant melanoma tissue.

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Linsler, S., A. Moroldo, J. Oertel, and S. Urbschat. "P18.05.A THE GROWTH PATTERN OF MENINGIOMA CELLS IN CELL CULTURE UNDER DIFFERENT CONDITIONS." Neuro-Oncology 25, Supplement_2 (September 1, 2023): ii122—ii123. http://dx.doi.org/10.1093/neuonc/noad137.413.

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Abstract BACKGROUND Cell cultures is an established method in tumor research. Thereby, the growth rate is important for the results. Depending on the experimental and clinical setting, the used tissue is exposed to different conditions. Interestingely, the different cell and tissue conditions are not well reported yet. Although, it is likely that the different conditions influence the growth pattern of cell cultures in primary cell lines. Here, the authors present their experimental analysis of cell culture of primary meningioma cells after different initial tissue conditions. MATERIAL AND METHODS From June to December 2022 ten primary human meningiomas were collected after surgery at the Neurosurgical Department of the Saarland University. Primary cell cultures (Passage 0 = P0) were set up on the day of surgery and one day postoperatively. These tissues were kept overnight in nutrient solution in the refrigerator. They were splitted in two P1 cultures (Passage 1). The termination of the cell culture occured when the flasks were fully grown. One P1 culture was used for Fluorescence in situ hybridization. The other one was frozen on liquid nitrogen. The latter is thawed after six months and cultivated again. Additionally, swab preparations were made for Fluorescence in situ hybridization. The frozen tissue of these meningiomas was also used for further cell cultures. RESULTS Eight of ten cases (80%) of the meningioma cell cultures (from the day of surgery and one day postoperatively) have grown. In one case only the cell culture from the day of surgery has grown and in another one there were only single cells, no growth out of this tissue. The cell cultures prepared from frozen tissues have not shown any culture growth. There were not any significant differences between the growth of the cell culture from the day of surgery and of the cell culture from one day after surgery. The results of the Fluorescence in situ hybridization have shown a loss of 1p in one meningioma, a loss of 22q in three meningiomas, a loss of 1p and 22q in three meningiomas and a normal chromosome set in three tumours. Further, there were not any differences between the evaluation of swab preparations and drop preparations. Until now, six out of six thawed cell cultures of the first three meningiomas can be cultured again after six months in liquid nitrogen. CONCLUSION It has been shown that the growth pattern of meningioma cell cultures is independent of the set up at the same day or one day after surgery if there are enough living cells in the tissue. On the other hand, meningioma cell culturing of frozen tissues is not possible in the same setting.

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6

Prakash, Jitendra. "Plant Tissue Culture." Nature Biotechnology 9, no. 7 (July 1991): 607. http://dx.doi.org/10.1038/nbt0791-607.

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7

Ezzell, Carol. "Tissue culture tools." Nature 327, no. 6119 (May 1987): 256–58. http://dx.doi.org/10.1038/327256a0.

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8

Prince, Cary. "Tissue culture trends." Nature 339, no. 6224 (June 1989): 488–90. http://dx.doi.org/10.1038/339488a0.

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Ezzell, Carol. "Tissue culture tips." Nature 333, no. 6173 (June 1988): 580–82. http://dx.doi.org/10.1038/333580a0.

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10

de Fossard, Ronald A. "Tissue-culture guide." Trends in Plant Science 6, no. 2 (February 2001): 85. http://dx.doi.org/10.1016/s1360-1385(00)01856-2.

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Dagla, H. R. "Plant tissue culture." Resonance 17, no. 8 (August 2012): 759–67. http://dx.doi.org/10.1007/s12045-012-0086-8.

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12

Seil, Fredrick J. "Tissue Culture Models of Myelination After Oligodendrocyte Transplantation." Journal of Neural Transplantation 1, no. 2 (1989): 49–55. http://dx.doi.org/10.1155/np.1989.49.

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Studies of myelination after transplantation of mature oligodendrocytes to cerebellar cultures in which oligodendrocyte maturation and myelination had been irreversibly inhibited by exposure to cytosine arabinoside were reviewed. Transplanted oligodendrocytes were derived from three sources, including cerebellar explants treated with kainic acid, dissociated oligodendrocyte cultures, and optic nerve fragments. Oligodendrocytes from all sources migrated into the host explants and myelinated appropriate axons. The time of appearance of myelin and the percentage of host cultures myelinated differed for the three sources of oligodendrocytes, however. Myelin was visible earliest and in the highest percentage of host explants transplanted with cultured dissociated oligodendrocytes, which were presumably the most free to migrate into the host tissue, and latest and in the lowest percentage of host cultures transplanted with optic nerve, from which oligodendrocytes were presumably least free to migrate. Some myelin-like membranes unassociated with axons appeared in cerebellar cultures transplanted with cultured dissociated oligodendrocytes, and not in cerebellar explants transplanted with oligodendrocytes from other sources. The formation of such myelin-like membranes was interpreted as a manifestation of oligodendrocyte hyperreactivity induced by culture in isolation.

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Stokes, Rebecca A., Michelle C. Coleman, Artem S. Rogovskyy, Vanna M. Dickerson, and Kelley M. Thieman Mankin. "Comparison of bacteriologic culture results for skin wound swabs and skin wound biopsy specimens." Journal of the American Veterinary Medical Association 259, no. 12 (December 15, 2021): 1416–21. http://dx.doi.org/10.2460/javma.20.10.0568.

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Abstract OBJECTIVE To compare bacteriologic culture results for superficial swab and tissue biopsy specimens obtained from dogs with open skin wounds. ANIMALS 52 client-owned dogs. PROCEDURES For each dog, 1 wound underwent routine preparation prior to collection of 2 specimens, 1 by superficial swab (Levine) technique and 1 by tissue biopsy. Specimens were processed for bacteriologic culture. Two observers determined whether any detected difference in culture results for the 2 types of specimen would have resulted in differing treatment plans. RESULTS Culture results of swab and tissue biopsy specimens were identical in 11/52 (21.2%) cases. Tissue biopsy specimen and swab cultures yielded positive results for 44 (84.6%) and 40 (76.9%) wounds, respectively. With regard to mean recovery rates of bacteria from wounds with positive culture results, both the biopsy specimens and swabs yielded 3.4 bacterial species/wound. All wounds for which swab cultures yielded no growth also had negative culture results for biopsy specimens. Biopsy specimen and swab culture results were in agreement with regard to the most common bacteria cultured. In 7/52 (13%) wounds, the observers would have treated the patient differently on the basis of the results of the 2 cultures. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that culture of a swab collected by the Levine technique is an appropriate noninvasive alternative to culture of a tissue biopsy specimen. A negative result obtained from culture of a swab is likely to be reliable. Disagreement between the results of swab and tissue biopsy specimen cultures is likely of low clinical importance.

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Sonwane, Ritesh, Rupesh Patil, Ketan Sonwane, Bhavesh Raghuwanshi, and Pragati Lokhande. "Tissue Culture: Tissue Plants Booking Website." Advancement of IoT in Blockchain Technology and its Applications 1, no. 1 (April 27, 2022): 8–10. http://dx.doi.org/10.46610/aibtia.2022.v01i01.002.

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Tissue Culture is an website for benefit of the farming community .Where farmers can book Tissue Plants at anytime from anywhere in this paper we discuss about a Website, from where Farmers can easily Book Plants like Banana, Papaya, strawberry, Pine apple etc. at cheap price and take higher yield form their crops.

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Cheng, Ya-Fu, Ching-Yuan Cheng, Chang-Lun Huang, Wei-Heng Hung, and Bing-Yen Wang. "Pleural Peels Tissue Culture plus Pleural Fluid Culture Help to Improve Culture Rate for Empyema." Journal of Clinical Medicine 11, no. 7 (March 28, 2022): 1882. http://dx.doi.org/10.3390/jcm11071882.

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Background: Empyema is known as a serious infection, and outcomes of empyema cases remain poor. Pleural fluid culture and blood culture have been reported to give unsatisfactory results. We introduce a novel pleural peels tissue culture during surgery and aim to improve the culture results of empyema. Methods: This was a retrospective study and was obtained from our institute. Patients with stage II or III empyema undergoing video-assisted thoracic surgery decortication from January 2019 to June 2021 were included in the study. Results: There were 239 patients that received a pleural peels tissue culture, a pleural fluid culture, and a blood culture concurrently during the perioperative period. Of these, 153 patients had at least one positive culture and 86 patients showed triple negative culture results. The positive culture rates were 46.9% for pleural peels tissue cultures, 46.0% for pleural fluid cultures, and 10% for blood cultures. The combination of pleural peels tissue culture and pleural fluid culture increased the positive rate to 62.7%. Streptococcus species and Staphylococcus species were the most common pathogens. Conclusion: The combination of pleural peels tissue culture and pleural fluid culture is an effective method to improve the positive culture rate in empyema.

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Tanini, Annalisa, Maria Luisa Brandi, Umberto Modigliani, Carlo M. Rotella, and Roberto Toccafondi. "Transient lack of response to TSH of human cultured thyroid cells obtained from hyperfunctioning tissue." Acta Endocrinologica 113, no. 3 (November 1986): 346–54. http://dx.doi.org/10.1530/acta.0.1130346.

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Abstract. TSH-induced cAMP accumulation in cells obtained from normal and pathological thyroid tissue was studied during the first 12 days of primary culture. In normal thyroid tissue cultures (N = 7), the response of cAMP to TSH was present from the second day of culture and reached its maximum after 8 days. A similar behaviour was observed in cultures obtained from euthyroid sporadic goitres (N = 8), even if the rate of response was slightly lower than that of normal tissue. Similarly, cultured cells from euthyroid 'autonomous' nodules (N = 8) appeared to be responsive to TSH during the period of study, but the rate of response was also lower than in the controls. On the contrary, in cultures obtained from toxic adenomas (N = 5) and from diffuse toxic goitres (N = 5) the response to TSH was absent during the first 4 days of culture. The cells became sensitive to TSH from 6 and 6 day onwards, with the rate of response increasing progressively and reaching its maximum on day 12. Finally, in cultured cells obtained from different areas of multinodular toxic goitres (N = 4), the response to TSH was similar to that of euthyroid goitres in cells prepared from 'cold' areas, and to that of toxic adenomas in cells obtained from 'hot' areas. The present data demonstrate the existence of an inhibitory action of unknown factors, possibly iodothyronines or thyroglobulin, on the TSH effect in short-term cultures obtained from thyrotoxic tissues. A normal TSH responsiveness can be restored when the culture is prolonged.

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Imashiro, Chikahiro, Kai Yamasaki, Ryu-ichiro Tanaka, Yusuke Tobe, Katsuhisa Sakaguchi, and Tatsuya Shimizu. "Perfusable System Using Porous Collagen Gel Scaffold Actively Provides Fresh Culture Media to a Cultured 3D Tissue." International Journal of Molecular Sciences 22, no. 13 (June 24, 2021): 6780. http://dx.doi.org/10.3390/ijms22136780.

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Culturing three-dimensional (3D) tissues with an appropriate microenvironment is a critical and fundamental technology in broad areas of cutting-edge bioengineering research. In addition, many technologies have engineered tissue functions. However, an effective system for transporting nutrients, waste, or oxygen to affect the functions of cell tissues has not been reported. In this study, we introduce a novel system that employs diffusion and convection to enhance transportation. To demonstrate the concept of the proposed system, three layers of normal human dermal fibroblast cell sheets are used as a model tissue, which is cultured on a general dish or porous collagen scaffold with perfusable channels for three days with and without the perfusion of culture media in the scaffold. The results show that the viability of the cell tissue was improved by the developed system. Furthermore, glucose consumption, lactate production, and oxygen transport to the tissues were increased, which might improve the viability of tissues. However, mechanical stress in the proposed system did not cause damage or unintentional functional changes in the cultured tissue. We believe that the introduced culturing system potentially suggests a novel standard for 3D cell cultures.

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Mothersill, Carmel, Colin Seymour, M. J. Moriarty, and M. J. Cullen. "Long-term culture of differentiated human thyroid tissue." Acta Endocrinologica 108, no. 2 (February 1985): 192–99. http://dx.doi.org/10.1530/acta.0.1080192.

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Abstract. Human thyroid cells obtained during surgery have been maintained in monolayer culture for at least 2 months and without loss of morphological or functional differentiation. Samples as small as 0.5 g could be cultured but best results were obtained with samples of 5–10 g. The technique used was developed in this laboratory for sheep tissue and was applicable without significant modification to human tissue. It depends on the complete absence of media changes at any time during the culture period. Energy substrates are replenished by the addition of concentrated glucose solutions to the existing media at carefully monitored intervals. Differences in both morphology and function could be observed between cultures derived from patients with different diseases, suggesting that the technique could have predictive value.

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Okano, T. "Muscular tissue engineering: capillary-incorporated hybrid muscular tissues in vivo tissue culture." Cell Transplantation 7, no. 5 (September 10, 1998): 435–42. http://dx.doi.org/10.1016/s0963-6897(98)00030-x.

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Okano, Takahisa, and Takehisa Matsuda. "Muscular Tissue Engineering: Capillary-Incorporated Hybrid Muscular Tissues in Vivo Tissue Culture." Cell Transplantation 7, no. 5 (September 1998): 435–42. http://dx.doi.org/10.1177/096368979800700502.

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Requirements for a functional hybrid muscular tissue are 1) a high density of multinucleated cells, 2) a high degree of cellular orientation, and 3) the presence of a capillary network in the hybrid tissue. Rod-shaped hybrid muscular tissues composed of C2C12 cells (skeletal muscle myoblast cell line) and type I collagen, which were prepared using the centrifugal cell-packing method reported in our previous article, were implanted into nude mice. The grafts, comprised three hybrid tissues (each dimension, diameter, approximately 0.3 mm, length, approximately 1 mm, respectively), were inserted into the subcutaneous spaces on the backs of nude mice. All nude mice that survived the implantation were sacrificed at 1, 2, and 4 wk after the implantation. The grafts were easily distinguishable from the subcutaneous tissues of host mice with implantation time. The grafts increased in size with time after implantation, and capillary networks were formed in the vicinities and on the surfaces of the grafts. One week after implantation, many capillaries formed in the vicinities of the grafts. In the central portion of the graft, few capillaries and necrotic cells were observed. Mononucleated myoblasts were densely distributed and a low number of multinucleated myotubes were scattered. Two weeks after implantation, the formation of a capillary network was induced, resulting in the surfaces of the grafts being covered by capillaries. Numerous elongated multinucleated myotubes and mononucleated myoblasts were densely distributed and numerous capillaries were observed throughout the grafts. Four weeks after implantation a dense capillary network was formed in the vicinities and on the surfaces of the grafts. In the peripheral portion of the graft, multinucleated myotubes in the vicinities of the rich capillaries were observed. Thus, hybrid muscular tissues in vitro preconstructed was remodeled in vivo, which resulted in facilitating the incorporation of capillary networks into the tissues. © 1998 Elsevier Science Inc.

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Peel, Trisha N., John A. Sedarski, Brenda L. Dylla, Samantha K. Shannon, Fazlollaah Amirahmadi, John G. Hughes, Allen C. Cheng, and Robin Patel. "Laboratory Workflow Analysis of Culture of Periprosthetic Tissues in Blood Culture Bottles." Journal of Clinical Microbiology 55, no. 9 (July 12, 2017): 2817–26. http://dx.doi.org/10.1128/jcm.00652-17.

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ABSTRACTCulture of periprosthetic tissue specimens in blood culture bottles is more sensitive than conventional techniques, but the impact on laboratory workflow has yet to be addressed. Herein, we examined the impact of culture of periprosthetic tissues in blood culture bottles on laboratory workflow and cost. The workflow was process mapped, decision tree models were constructed using probabilities of positive and negative cultures drawn from our published study (T. N. Peel, B. L. Dylla, J. G. Hughes, D. T. Lynch, K. E. Greenwood-Quaintance, A. C. Cheng, J. N. Mandrekar, and R. Patel, mBio 7:e01776-15, 2016,https://doi.org/10.1128/mBio.01776-15), and the processing times and resource costs from the laboratory staff time viewpoint were used to compare periprosthetic tissues culture processes using conventional techniques with culture in blood culture bottles. Sensitivity analysis was performed using various rates of positive cultures. Annualized labor savings were estimated based on salary costs from the U.S. Labor Bureau for Laboratory staff. The model demonstrated a 60.1% reduction in mean total staff time with the adoption of tissue inoculation into blood culture bottles compared to conventional techniques (mean ± standard deviation, 30.7 ± 27.6 versus 77.0 ± 35.3 h per month, respectively;P< 0.001). The estimated annualized labor cost savings of culture using blood culture bottles was $10,876.83 (±$337.16). Sensitivity analysis was performed using various rates of culture positivity (5 to 50%). Culture in blood culture bottles was cost-effective, based on the estimated labor cost savings of $2,132.71 for each percent increase in test accuracy. In conclusion, culture of periprosthetic tissue in blood culture bottles is not only more accurate than but is also cost-saving compared to conventional culture methods.

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Potts, Robert W. A., Alejandro P. Gutierrez, Yennifer Cortés-Araya, Ross D. Houston, and Tim P. Bean. "Developments in marine invertebrate primary culture reveal novel cell morphologies in the model bivalve Crassostrea gigas." PeerJ 8 (June 1, 2020): e9180. http://dx.doi.org/10.7717/peerj.9180.

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Cell culture provides useful model systems used in a wide range of biological applications, but its utility in marine invertebrates is limited due to the lack of immortalised cell lines. Primary cell and tissue cultures are typically used but remain poorly characterised for oysters, which can cause issues with experimental consistency and reproducibility. Improvements to methods of repeatable isolation, culture, and characterisation of oyster cells and tissues are required to help address these issues. In the current study, systematic improvements have been developed to facilitate the culture of primary cells from adult Pacific oyster tissues and identify novel cell morphologies that have not been reported previously. Cultures analysed by light microscopy, qPCR, and live cell imaging demonstrated maintenance of live, metabolically active Pacific oyster cells for several weeks post-explant. Interestingly, whole hearts dissected from adult oysters were found to continue contracting rhythmically up to 8 weeks after being transferred to a tissue culture system. Mantle tissue explants were also actively moving in the culture system. These improvements in primary cell culture of bivalves may be beneficial for research in ecotoxicology, virology, immunology, and genetic resistance to disease.

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Ogita, Shinjiro, Junko Miyazaki, Toshinari Godo, and Yasuo Kato. "Possibility for Selective Accumulation of Polyphenolics in Tissue Cultures of Senno (Lychnis Senno Siebold et Zucc.)." Natural Product Communications 4, no. 3 (March 2009): 1934578X0900400. http://dx.doi.org/10.1177/1934578x0900400312.

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Senno ( Lychnis senno Siebold et Zucc.), a traditional ornamental plant in Japan had been used as a crude drug acting as natural blood thinners. Since tissue culture protocols have been established, we analyzed polyphenol accumulation profiles in shoot culture, multiple shoot culture, and callus culture using the technique of HPLC with a Photodiode Array Detector. By comparing the HPLC profiles at 220-400 nm from extracts of different cultures, 14 putative flavonoids were confirmed as major metabolites in the cultures of Senno. Among the 14 compounds detected, 6 were tissue specific metabolites. It appears that the biosynthetic pathway of polyphenolics in Senno is regulated or strongly influenced by how tissues are regenerated and maintained in the in vitro environment. Hence, it may be possible to selectively produce novel secondary metabolites including flavonoids by engineering a target tissue culture procedure developed in the present study.

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Vogt, P. M., F. W. Peter, and H. U. Steinau. "Perspectives of tissue transfer and tissue culture." Der Orthopäde 27, no. 1 (February 6, 1998): 45–50. http://dx.doi.org/10.1007/pl00003449.

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Dey, Nandini, Yuliang Sun, Amy K. Krie, Luis Rojas, David Starks, Xiaoqian Lin, Kirstin Anne Williams, et al. "Three dimensional organotypic ex vivo culture of tissues from post-operated tumor samples: Strengths and limitations." Journal of Clinical Oncology 35, no. 15_suppl (May 20, 2017): e23151-e23151. http://dx.doi.org/10.1200/jco.2017.35.15_suppl.e23151.

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e23151 Background: Evolution of tumor occurs in two phases, pretreatment phase, and posttreatment phase.Tumorigenic history and path of tumorigenic evolution determine the response of tumor cells to antitumor drug and thus the outcome of treatment. Carcinomas from the same organ-site with similar genomic alterations in different patients may vary in their tumorigenic history and their diverse paths of tumorigenic evolution which cause them respond differently to the same antitumor drugs. Uniqueness of tumorigenic history and paths of tumorigenic evolution in individual patient makes it ideal to test drugs, N-of-1. Here we present our experience with 3D organotypic ex vivo culture of tumor tissue from post-operated tumor samples. Methods: Informed consent was obtained from patients ( NCT02470715). The investigation was approved by IRB of the Avera Cancer Institute and the Western IRB. Surgically resected tissue was obtained from patients with different tumors of different organ sites including ovarian, breast, lung, endometrium, and cervix at the Avera Cancer Institute, SF, SD. Tissues from patients were collected in two formats, paired samples (tumor and tumor-adjacent normal) and unpaired samples. Results: As extracellular matrix lost in cell cultures, provide important signals for cell survival, differentiation and drug resistance we have established a protocol to culture slices in the 3D format. We have established 3D slice cultures (3x3 mm) of tumor-derived tissues which can be maintained ex vivo for at least three days. Tumor cells / normal cells from different tumors/tumor-adjacent normal tissues from day zero (non-cultured) were used to compare day1, day2, and day3 cultures. At the end of the culture period, slices were embedded in paraffin for histological analysis following H&E staining as well as IHC-staining for proliferation index (Ki-67), cellular proliferation signals (pS6RP), apoptosis (activated caspase 3), or tumor angiogenesis (CD31). Ki67 staining was characteristically different in cultured normal and cultured tumor tissues. Conclusions: Here, we show that tumor-derived tissues survive in 3D Matrigel culture for a minimum of 6 hours and maximum up to 3 days. Clinical trial information: NCT02470715.

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26

Beck, K. L., J. Singh, and M. Anzar. "47 THE AVIAN CHORIO-ALLANTOIC MEMBRANE: A SUITABLE SHORT-TERM CULTURE SYSTEM FOR BOVINE OVARIAN TISSUE." Reproduction, Fertility and Development 27, no. 1 (2015): 116. http://dx.doi.org/10.1071/rdv27n1ab47.

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Successful cryopreservation of bovine ovarian tissue holds enormous potential for long-term maintenance of female gametes to preserve genetic diversity by tissue banking. Traditionally, in vitro culture followed by histopathological examination has been used to assess the post-thaw viability of cryopreserved tissues. Recently, in ovo transplantation of mammalian tissues on the chorio-allantoic membrane (CAM) of a growing chicken embryo has emerged as an alternative method for short-term culture. The purpose of this experiment was to compare CAM culture of bovine ovarian tissue over a 5-day period with the in vitro culture system. Fertilized White Leghorn eggs were incubated at 37°C and 62% relative humidity. A window (1 × 2 cm) was cut into the eggshell on Day 3 of incubation. Ovaries were retrieved from a local abattoir and brought to the laboratory within 6 h. Ovarian cortex pieces (1–2 mm3) were randomly assigned to control, CAM-culture, or in vitro-culture groups. Control-group tissues were fixed immediately in 4% paraformaldehyde. The CAM was traumatized on Day 10 of incubation to expose the underlying blood vessels, and tissue pieces were grafted at the site (one graft per egg). For in vitro culture, the ovarian cortex pieces were placed on tissue culture inserts within 6-well plates containing TCM199 with 1% insulin-transferrin-selenium, 100 mIU mL–1 of FSH, 100 IU mL–1 of penicillin, and 50 μg mL–1 of streptomycin and incubated at 38°C in 5% CO2. Ovarian tissues from the CAM and in vitro culture group were removed on Day 1, 3, and 5 of grafting/culture, fixed, embedded in paraffin, sectioned at 5 μm, stained with hematoxylin-eosin, and analysed under a light microscope. The numbers of normal and degenerated follicles (indicating follicle survival) and number of blood vessels containing bovine and avian red blood cells (indicating angiogenesis) were counted using standard stereological procedures. All ovarian cortex grafts from surviving chick embryos showed adhesion with the CAM and a marked neo-vascularization in the graft areas. Gross and histological examination revealed the circulation of avian blood cells in ovarian stromal vessels with a concomitant decrease in the number of bovine blood vessels over the incubation period. Total follicle densities (mean ± s.e.m.) on Day 1, 3, and 5 were 13.3 ± 5.9, 27.9 ± 6.7, and 36.9 ± 7.3 in the in vitro-cultured group and 36.7 ± 13.0, 73.6 ± 24.0, and 44.02 ± 12.67 per millimeter cubed in the CAM-cultured group, respectively. Overall, total follicle density was higher in the CAM-cultured group (P < 0.05). Likewise, the normal follicle densities on Day 1, 3, and 5 were 10.4 ± 4.9, 15.5 ± 3.6, and 20.7 ± 6.3 in the in vitro-cultured group and 30.5 ± 8.5, 45.7 ± 18.4, and 22.7 ± 7.3 per millimeter cubed in the CAM-cultured group (P > 0.05). In conclusion, in ovo CAM grafting system was as successful as the in vitro-culture system and may be considered an acceptable alternative to the traditional in vitro-culture system for bovine ovarian tissue.

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Kreß, Sebastian, Roland Schaller-Ammann, Jürgen Feiel, Joachim Wegener, Joachim Priedl, Wolf Dietrich, Cornelia Kasper, and Dominik Egger. "Innovative Platform for the Advanced Online Monitoring of Three-Dimensional Cells and Tissue Cultures." Cells 11, no. 3 (January 25, 2022): 412. http://dx.doi.org/10.3390/cells11030412.

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The use of 3D cell cultures has gained increasing importance in medical and pharmaceutical research. However, the analysis of the culture medium is hardly representative for the culture conditions within a 3D model which hinders the standardization of 3D cultures and translation of results. Therefore, we developed a modular monitoring platform combining a perfusion bioreactor with an integrated minimally invasive sampling system and implemented sensors that enables the online monitoring of culture parameters and medium compounds within 3D cultures. As a proof-of-concept, primary cells as well as cell lines were cultured on a collagen or gelatin methacryloyl (GelMA) hydrogel matrix, while monitoring relevant culture parameters and analytes. Comparing the interstitial fluid of the 3D models versus the corresponding culture medium, we found considerable differences in the concentrations of several analytes. These results clearly demonstrate that analyses of the culture medium only are not relevant for the development of standardized 3D culture processes. The presented bioreactor with an integrated sampling and sensor platform opens new horizons for the development, optimization, and standardization of 3D cultures. Furthermore, this technology holds the potential to reduce animal studies and improve the transferability of pharmaceutical in vitro studies by gaining more relevant results, bridging the gap towards clinical translation.

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Encina, C. L., I. M. G. Padilla, J. M. Cazorla, and E. Caro. "TISSUE CULTURE IN CHERIMOYA." Acta Horticulturae, no. 497 (August 1999): 289–302. http://dx.doi.org/10.17660/actahortic.1999.497.15.

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YAMAMOTO, Yoshikazu. "Tissue Culture of Lichens." Plant tissue culture letters 5, no. 1 (1988): 44–46. http://dx.doi.org/10.5511/plantbiotechnology1984.5.44.

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Loeb, Marcia, and Jun Mitsuhashi. "Invertebrate Tissue Culture Methods." Annals of the Entomological Society of America 96, no. 3 (May 1, 2003): 344. http://dx.doi.org/10.1603/0013-8746(2003)096[0344:itcm]2.0.co;2.

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31

Illg, Rolf Dieter. "Plant tissue culture techniques." Memórias do Instituto Oswaldo Cruz 86, suppl 2 (1991): 21–24. http://dx.doi.org/10.1590/s0074-02761991000600008.

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32

HORTON, BRENDAN. "Cell and tissue culture." Nature 381, no. 6579 (May 1996): 255–58. http://dx.doi.org/10.1038/381255a0.

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33

Ruben, Regina L. "The new tissue culture." Science 356, no. 6335 (April 20, 2017): 342. http://dx.doi.org/10.1126/science.356.6335.342.

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Hickey, Ivor. "Tissue culture without tears." Trends in Cell Biology 3, no. 12 (December 1993): 453. http://dx.doi.org/10.1016/0962-8924(93)90042-y.

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35

Messenger, A. G. "Hair follicle tissue culture." British Journal of Dermatology 113, no. 6 (December 1985): 639–40. http://dx.doi.org/10.1111/j.1365-2133.1985.tb02397.x.

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Poginsky, B., W. Preil, J. Westendorf, and Lj Kraus. "Tissue Culture ofRubia tinctorum." Planta Medica 55, no. 02 (April 1989): 231–32. http://dx.doi.org/10.1055/s-2006-961989.

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37

J. Robins, Ichard. "Plant tissue culture manual." Phytochemistry 31, no. 9 (September 1992): 3301–2. http://dx.doi.org/10.1016/0031-9422(92)83507-u.

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Xiang, Yu, Jiongyi Yan, Xujin Bao, Andrew Gleadall, Paul Roach, and Tao Sun. "Evaluation of Polymeric Particles for Modular Tissue Cultures in Developmental Engineering." International Journal of Molecular Sciences 24, no. 6 (March 9, 2023): 5234. http://dx.doi.org/10.3390/ijms24065234.

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Developmental engineering (DE) aims to culture mammalian cells on corresponding modular scaffolds (scale: micron to millimeter), then assemble these into functional tissues imitating natural developmental biology processes. This research intended to investigate the influences of polymeric particles on modular tissue cultures. When poly(methyl methacrylate) (PMMA), poly(lactic acid) (PLA) and polystyrene (PS) particles (diameter: 5–100 µm) were fabricated and submerged in culture medium in tissue culture plastics (TCPs) for modular tissue cultures, the majority of adjacent PMMA, some PLA but no PS particles aggregated. Human dermal fibroblasts (HDFs) could be directly seeded onto large (diameter: 30–100 µm) PMMA particles, but not small (diameter: 5–20 µm) PMMA, nor all the PLA and PS particles. During tissue cultures, HDFs migrated from the TCPs surfaces onto all the particles, while the clustered PMMA or PLA particles were colonized by HDFs into modular tissues with varying sizes. Further comparisons revealed that HDFs utilized the same cell bridging and stacking strategies to colonize single or clustered polymeric particles, and the finely controlled open pores, corners and gaps on 3D-printed PLA discs. These observed cell–scaffold interactions, which were then used to evaluate the adaptation of microcarrier-based cell expansion technologies for modular tissue manufacturing in DE.

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Winkelkotte, Maximilian, Florian Schmieder, Stephan Behrens, Dominic Salminger, Anett Jannasch, Klaus Matschke, Sems Malte Tugtekin, Frank Sonntag, and Claudia Dittfeld. "Micro-Physiological-Systems enable investigation of hypoxia induced pathological processes in human aortic valve cells and tissues." Current Directions in Biomedical Engineering 7, no. 2 (October 1, 2021): 45–48. http://dx.doi.org/10.1515/cdbme-2021-2012.

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Abstract Aortic valve (AV) stenosis is characterized by tissue fibrosis and calcification. Fibrous thickening can result in reduced tissue oxygen supply leading to pathological valvular interstitial cell (VIC) differentiation and calcification. Static 2D VIC cultures and animal models are limited in the ability to reflect human AV calcification. Culturing of VICs in micro-physiological-systems (MPS) in a pulsatile flow and the establishment of a modular AV tissue incubation chamber (TIC) are new approaches to evaluate pathophysiological processes of AV disease. Therefore, a MPS able to adjust hypoxic conditions was applied for VIC culture. A significant increase of mRNA-expression of EGLN1 and HIF1α- regulated LDHA and HIF1α nuclear localisation were proven under hypoxia. AV tissue culture was established within a TIC and viability was monitored by Resazurin-reduction in the incubation medium and visualized by LDH-activity in tissue cryosections. Viability was compared between fluid and static incubated tissues revealing an advantageous effect of the fluidic assay condition. Consecutively, the application of MPS in AV research allows i) the investigation of VIC cultures with efficient oxygen regulation and ii) the culture of porcine or human AV tissues preserving viability and specifically reflecting in vivo parameters. These methods open up new possibilities beyond static 2D culture and facilitate a reduction of animal experiments in AV research.

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Anushi, Shubham Jain, Manjunath Rathod, Gopa Mishra, V. Lakshmi Prasanna Kumari, Hari Baksh, Saransh Saxena, and Lalu Prasad. "Plant Tissue Culture for Medical Therapy: Unlocking the Potential of Medicinal Plants." Current Journal of Applied Science and Technology 42, no. 46 (December 1, 2023): 7–22. http://dx.doi.org/10.9734/cjast/2023/v42i464289.

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Plant tissue culture is emerging as a pivotal biotechnological tool with profound implications for medical therapy, particularly in the realm of herbal medicine. Medicinal plants have long been cherished for their natural healing properties. However, escalating demand, habitat destruction, and overharvesting have threatened the availability and sustainability of these valuable resources. Plant tissue culture addresses these concerns by enabling the mass propagation of medicinal plants. In controlled environments, plant tissues can be multiplied rapidly, providing a continuous and sustainable source of plant material. This not only safeguards wild populations but also ensures a consistent supply of bioactive compounds that form the basis of herbal therapies. One of the most transformative applications of plant tissue culture in medical therapy is the manipulation of secondary metabolite production. Medicinal plants synthesize a diverse array of bioactive compounds, such as alkaloids, flavonoids, and terpenoids, with therapeutic properties. Through precise control of growth conditions and genetic modification, plant tissue culture can enhance the yield of these compounds, thereby increasing the potency and efficacy of herbal medicines. This precision is instrumental in the pharmaceutical industry, where the isolation and production of specific bioactive compounds can lead to the development of novel drugs and therapies. In addition to bolstering yields, plant tissue culture offers the advantage of disease-free plant material. By maintaining cultures in sterile conditions, the risk of contaminants and pathogens is mitigated, enhancing the safety and quality of herbal medicines. These cultures can also serve as a continuous source of plant-derived compounds, enabling a consistent supply of bioactive substances. Furthermore, plant tissue culture is a crucial tool for research and development in the field of medicinal plants. It provides a controlled platform for studying plant biology, optimizing growth conditions, and investigating the mechanisms underlying secondary metabolite production. These insights contribute to the development of improved plant varieties with enhanced medicinal properties, addressing the evolving needs of medical therapy. While the potential of plant tissue culture in medical therapy is vast, it is essential to underscore the importance of rigorous research, quality control, and safety assessments. Ensuring the safety and efficacy of products derived from tissue-cultured plants is paramount to their acceptance and use in medical applications. Compliance with regulatory standards and collaboration with healthcare professionals are integral to upholding the quality and safety of medicinal products.

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Twardovska, M. O. "THE CONTENT OF PHENOLIC COMPOUNDS AND FLAVONOIDS IN Deschampsia antarctica TISSUE CULTURE." Biotechnologia Acta 14, no. 2 (February 2021): 59–66. http://dx.doi.org/10.15407/biotech14.02.059.

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Aim. The aim of the study was to determine the quantitative and qualitative content of phenolic compounds and flavonoids in Deschampsia antarctica E. Desv. tissue cultures obtained from plants originating from different islands of the maritime Antarctic. Methods. In vitro tissue culture, Folin-Ciocalteu method, spectrophotometry, HPLC analysis. Results. The quantitative content of phenolic compounds and flavonoids in D. antarctica tissue cultures obtained from plants of six genotypes (DAR12, DAR13, G/D12-2a, Y66, R30 and L57) was determined. The highest content of phenolic compounds (4.46 and 3.75 mg/g) was found in tissue cultures obtained from root and leaf explants of plant genotype L57. The highest amount of flavonoids (7.17 mg/g) was accumulated in G/D12-2a tissue culture of root origin. The content of the studied biologically active compounds (BACs) did not change with increasing number of subculture generations (from passage 10 to 19). HPLC analysis showed that in D. antarctica tissue cultures, a shift in the biosynthesis of BACs occurred towards the synthesis of more polar metabolites compared to explant donor plants. Conclusions. It was found that the transition of cells to undifferentiated growth affected the content of BACs, the amount of which decreased 2–5 times simultaneously with a significant change in their profile. This provided a basis for further biochemical studies, as well as for careful selection of tissue culture of D. antarctica to use it as a potential source of BACs.

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Polisetti, Naresh, Gottfried Martin, Eva Ulrich, Mateusz Glegola, Ursula Schlötzer-Schrehardt, Günther Schlunck, and Thomas Reinhard. "Influence of Organ Culture on the Characteristics of the Human Limbal Stem Cell Niche." International Journal of Molecular Sciences 24, no. 23 (November 28, 2023): 16856. http://dx.doi.org/10.3390/ijms242316856.

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Organ culture storage techniques for corneoscleral limbal (CSL) tissue have improved the quality of corneas for transplantation and allow for longer storage times. Cultured limbal tissue has been used for stem cell transplantation to treat limbal stem cell deficiency (LSCD) as well as for research purposes to assess homeostasis mechanisms in the limbal stem cell niche. However, the effects of organ culture storage conditions on the quality of limbal niche components are less well described. Therefore, in this study, the morphological and immunohistochemical characteristics of organ-cultured limbal tissue are investigated and compared to fresh limbal tissues by means of light and electron microscopy. Organ-cultured limbal tissues showed signs of deterioration, such as edema, less pronounced basement membranes, and loss of the most superficial layers of the epithelium. In comparison to the fresh limbal epithelium, organ-cultured limbal epithelium showed signs of ongoing proliferative activity (more Ki-67+ cells) and exhibited an altered limbal epithelial phenotype with a loss of N-cadherin and desmoglein expression as well as a lack of precise staining patterns for cytokeratin ((CK)14, CK17/19, CK15). The analyzed extracellular matrix composition was mainly intact (collagen IV, fibronectin, laminin chains) except for Tenascin-C, whose expression was increased in organ-cultured limbal tissue. Nonetheless, the expression patterns of cell–matrix adhesion proteins varied in organ-cultured limbal tissue compared to fresh limbal tissue. A decrease in the number of melanocytes (Melan-A+ cells) and Langerhans cells (HLA-DR+, CD1a+, CD18+) was observed in the organ-cultured limbal tissue. The organ culture-induced alterations of the limbal epithelial stem cell niche might hamper its use in the treatment of LSCD as well as in research studies. In contrast, reduced numbers of donor-derived Langerhans cells seem associated with better clinical outcomes. However, there is a need to consider the preferential use of fresh CSL for limbal transplants and to look at ways of improving the limbal stem cell properties of stored CSL tissue.

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Das, Debarath, Kishore D, Lakshmana R, Pravin Dhas, Snigdha N, and Malarmannan M. "Comparative study of tissue culture and sensitivity versus swab culture and sensitivity of microorganisms in the healing of diabetic foot ulcers." International Journal of Research in Pharmaceutical Sciences 15, no. 1 (February 12, 2024): 25–31. http://dx.doi.org/10.26452/ijrps.v15i1.4660.

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This prospective observational study, conducted at the Department of General Surgery, SRM Medical College and Hospital, aimed to assess the effectiveness of tissue culture and sensitivity compared to swab culture and sensitivity in the healing of diabetic foot ulcers through antibiotic sensitivity of microorganisms. Between May 2016 and August 2017, 160 subjects with diabetic foot ulcers were randomly assigned treatment based on either swab or tissue culture findings. Patients were followed at 15-day intervals for up to 60 days. Results showed positive swab cultures in 76.88% and positive tissue cultures in 92.50% of the study population. The most prevalent organism in swab cultures was Proteus (14.38%), while Pseudomonas (16.88%) dominated in tissue cultures. The cumulative proportion of subjects developing granulation tissue was faster in the tissue culture group, reaching 57.50% at 15 to 30 days and 99% at 31 to 45 days. The swab culture group exhibited proportions of 48.80%, 75%, and 93.80% at the same intervals. In conclusion, diabetic foot ulcer treatment based on tissue culture showed slightly faster healing rates compared to swab culture. However, both groups achieved good ulcer healing within the 60-day follow-up period. These findings emphasize the importance of choosing an appropriate culture method for effective management of diabetic foot ulcers.

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B.M. Johri, P.S. Srivastava, and Madhumati Purohit. "Plant tissue culture and biotechnology." Journal of Palaeosciences 46, no. 3 (December 31, 1997): 134–59. http://dx.doi.org/10.54991/jop.1997.1357.

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Plant tissue culture has progressed steadily ever since its inception in 1902. The initial experiments related to various tissues that could sustain prolonged in vitro conditions. The differential response of the cultured tissues under variable chemical milieu provided the necessary impetus to utilize the technique in a profitable manner. Over the years efficacy of the technique became apparent when noticeable in vitro morphogenic responses could be used to unravel the mysteries of growth and differentiation. Expectedly, therefore, any morphogenic event expressed in vitro could be correlated to the specific components of the nutritive medium. By the 1970s the applicability of the technique came to be realized with the possibility of exploring somatic hybridization, micropropagation of recalcitrant species, haploid, and triploid plants, and finally genetic manipulations. Today, plant tissue culture has become an integral part of biotechnology and is being routinely employed for the improvement of crops and legumes- the backbone of human nutrition that can also aid in the amelioration of malnutrition of millions of sufferers. The ultimate success with the transfer of 'nif’-gene to non-leguminous plants would help save millions of dollars in chemical fertilizers which can then be profitably used for the welfare of the human race.

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Kunakh, V. A., D. O. Navrotska, M. O. Twardovska, and I. O. Andreev. "Peculiarities of chromosomal variability in cultured tissues of Deschampsia antarctica Desv. plants with different chromosome numbers." Visnik ukrains'kogo tovaristva genetikiv i selekcioneriv 14, no. 1 (June 20, 2016): 36–43. http://dx.doi.org/10.7124/visnyk.utgis.14.1.542.

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Aim. To clarify the details of chromosome variation in calli derived from D. antarctica plants in the initial passages of the culture in vitro. Methods. Induction of callus from root explants of plants, which were grown from seeds, and consequent subcultivation of tissue culture. Cytogenetic analysis of squashed slides stained by acetic-orcein and counting the number of chromosomes in mitotic metaphase plates. Results. There were analyzed the cultured tissues derived from D. antarctica plants with different chromosome numbers: diploid plants (2n=26), mixoploid plant with B-chromosomes (2n=26+1-3B), and mixoploid plant with near-triploid modal class (2n=36, 38). Analysis of callus tissues of all plants at 2-4 passages revealed mixoploidy, presence of polyploid and aneuploid cells. The modal class in all studied calli was composed of diploid and aneuploid cells with near-diploid chromosome number. The cytogenetic structure of cell population of cultured tissues was found to vary with characteristics of the karyotype of donor plant. The largest range of variation in the number of chromosomes (from 18 to 63 chromosomes) was found in tissue culture of diploid plant (2n=26) from the Galindez Island, and the highest frequencies of polyploid (47 %) and aneuploid cells were in the culture of mixoploid plant with near-triploid modal class from the Big Yalour Island. Conclusions. In different D. antarctica cultured tissues at the early stages of the culture, the modal class was composed of diploid cells and cells with near-diploid chromosome number irrespective of karyotype of donor plant (diploid, mixoploid poliploid).Key words: Deschampsia antarctica Desv., plant tissue culture, chromosomal variability in vitro, mixoploidy.

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Sahashi, Yuko, Naoki Yumiza, Koichi Kamekura, and Kazuo Sugaya. "Culture conditions for stable cultivation of tissue-cultured ginseng." Journal of Bioscience and Bioengineering 108 (November 2009): S5. http://dx.doi.org/10.1016/j.jbiosc.2009.08.022.

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47

Sudhersan, C., Y. Al-Shayji, S. Jibimanuel, and J. Ashkanani. "PHOTOAUTOTROPHIC CULTURE PHASE FOR TISSUE CULTURED DATE PALM PLANTLETS." Acta Horticulturae, no. 994 (June 2013): 313–21. http://dx.doi.org/10.17660/actahortic.2013.994.31.

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Martin-Iglesias, Sara, Lara Milian, María Sancho-Tello, Rubén Salvador-Clavell, José Javier Martín de Llano, Carmen Carda, and Manuel Mata. "BMP-2 Enhances Osteogenic Differentiation of Human Adipose-Derived and Dental Pulp Stem Cells in 2D and 3D In Vitro Models." Stem Cells International 2022 (March 4, 2022): 1–15. http://dx.doi.org/10.1155/2022/4910399.

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Bone tissue provides support and protection to different organs and tissues. Aging and different diseases can cause a decrease in the rate of bone regeneration or incomplete healing; thus, tissue-engineered substitutes can be an acceptable alternative to traditional therapies. In the present work, we have developed an in vitro osteogenic differentiation model based on mesenchymal stem cells (MSCs), to first analyse the influence of the culture media and the origin of the cells on the efficiency of this process and secondly to extrapolate it to a 3D environment to evaluate its possible application in bone regeneration therapies. Two osteogenic culture media were used (one commercial from Stemcell Technologies and a second supplemented with dexamethasone, ascorbic acid, glycerol-2-phosphate, and BMP-2), with human cells of a mesenchymal phenotype from two different origins: adipose tissue (hADSCs) and dental pulp (hDPSCs). The expression of osteogenic markers in 2D cultures was evaluated in several culture periods by means of the immunofluorescence technique and real-time gene expression analysis, taking as reference MG-63 cells of osteogenic origin. The same strategy was extrapolated to a 3D environment of polylactic acid (PLA), with a 3% alginate hydrogel. The expression of osteogenic markers was detected in both hADSCs and hDPSCs, cultured in either 2D or 3D environments. However, the osteogenic differentiation of MSCs was obtained based on the culture medium and the cell origin used, since higher osteogenic marker levels were found when hADSCs were cultured with medium supplemented with BMP-2. Furthermore, the 3D culture used was suitable for cell survival and osteogenic induction.

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Brückner, Luzi, Annika Reinshagen, Bahriye Aktas, Ingo Bechmann, Ivonne Nel, and Sonja Kallendrusch. "Development of a personalized ex vivo drug screening test for patients with ovarian cancer." Journal of Clinical Oncology 38, no. 15_suppl (May 20, 2020): e18090-e18090. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.e18090.

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e18090 Background: The standard therapy of patients with ovarian cancer consists of primary surgery followed by chemotherapy. Initial response rates are very high, but recurrence occurs in 85% of the cases. Personalized ex vivo analyses of various anti-tumor compounds in a standardized tissue slice culture system (1) might be a very promising approach for individualized therapeutic decisions. In comparison to cell culture, tumor slice cultures maintain the direct tumor microenvironment which plays a role in resistance mechanisms and thus therapy response. Methods: Patient derived tumor cultures (1) are grown under standardized conditions and are analyzed semi-automated. Patient’s tumor samples were collected during surgery, cut into standardized slices and were cultivated in triplicates for 2, 4, 7 and 14 days and treated with standard therapy for 7 days. A baseline control was prepared at day 0. The cultured tissue is PFA-fixated and paraffin embedded. Hematoxylin eosin staining was performed for microscopic evaluation of morphologic structures. Subsequently, immunohistochemical staining against CD3 was applied to examine the immune setting of the tumor and its environment. Results: Ovarian tumor tissues remained their morphological properties over a period of 14 days. Parameters like cellular formation, proliferation and heterogeneity were adequately represented in the cultures.. Staining against CD3 revealed T-cells in ovarian tumor tissue slices up to 14 days ex vivo and individual response to treatment was observable. Conclusions: Correlation to clinical data is ongoing to analyze the tissue culture model of ovarian cancer for clinical usage. Different approaches, concerning mutational burden, immunological signature and histology are considered for decision of response and non-response.

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Wightman, Raymond, and C. J. Luo. "From mammalian tissue engineering to 3D plant cell culture." Biochemist 38, no. 4 (August 1, 2016): 32–35. http://dx.doi.org/10.1042/bio03804032.

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Recent advances applying mammalian tissue engineering to in vitro plant cell culture have successfully cultured single plant cells in a 3D microstructure, leading to the discovery of plant cell behaviours that were previously not envisaged. Animal and plant cells share a number of properties that rely on a hierarchical microenvironment for creating complex tissues. Both mammalian tissue engineering and 3D plant culture employ tailored scaffolds that alter a cell's behaviour from the initial culture used for seeding. For humans, these techniques are revolutionizing healthcare strategies, particularly in regenerative medicine and cancer studies. For plants, we predict applications both in fundamental research to study morphogenesis and for synthetic biology in the agri-biotech sector.

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