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Journal articles on the topic 'Explant cultures'

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Author: Grafiati
Published: 10 December 2022
Last updated: 28 January 2023

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1

Žižková, Radmila, Věra Hedvičáková, Veronika Hefka Blahnová, Věra Sovková, Michala Rampichová, and Eva Filová. "The Effect of Osteoblast Isolation Methods from Adult Rats on Osteoclastogenesis in Co-Cultures." International Journal of Molecular Sciences 23, no. 14 (July 17, 2022): 7875. http://dx.doi.org/10.3390/ijms23147875.

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Co-cultures of osteoblasts and osteoclasts are on the rise because they enable a more complex study. Diseases such as osteoporosis are related to a higher age. Thus, cell isolation from adult individuals is necessary. Osteoblasts can be isolated from the rat femur by three methods: explant culture, explant culture with enzymatic pre-treatment, or enzymatic treatment. The isolation methods yield different populations of osteoblasts which, in a co-culture with peripheral blood mononuclear cells, might result in differences in osteoclastogenesis. Therefore, we examined the differences in osteogenic markers, cell proliferation, and the metabolic activity of isolated osteoblast-like cells in a growth and differentiation medium. We then evaluated the effect of the isolated populations of osteoblast-like cells on osteoclastogenesis in a subsequent co-culture by evaluating osteoclast markers, counting formed osteoclast-like cells, and analyzing their area and number of nuclei. Co-cultures were performed in the presence or absence of osteoclastogenic growth factors, M-CSF and RANKL. It was discovered that enzymatic isolation is not feasible in adult rats, but explant culture and explant culture with enzymatic pre-treatment were both successful. Explant culture with enzymatic pre-treatment yielded cells with a higher proliferation than explant culture in a growth medium. The differentiation medium reduced differences in proliferation during the culture. Some differences in metabolic activity and ALP activity were also found between the osteoblast-like cells isolated by explant culture or by explant culture with enzymatic pre-treatment, but only on some days of cultivation. According to microscopy, the presence of exogenous growth factors supporting osteoclastogenesis in co-cultures was necessary for the formation of osteoclast-like cells. In this case, the formation of a higher number of osteoclast-like cells with a larger area was observed in the co-culture with osteoblast-like cells isolated by explant culture compared to the explant culture with enzymatic pre-treatment. Apart from this observation, no differences in osteoclast markers were noted between the co-cultures with osteoblast-like cells isolated by explant culture and the explant culture with enzymatic pre-treatment. The TRAP and CA II activity was higher in the co-cultures with exogenous growth than that in the co-cultures without exogenous growth factors on day 7, but the opposite was true on day 14. To conclude, explant culture and explant culture with enzymatic pre-treatment are both suitable methods to yield osteoblast-like cells from adult rats capable of promoting osteoclastogenesis in a direct co-culture with peripheral blood mononuclear cells. Explant culture with enzymatic pre-treatment yielded cells with a higher proliferation. The explant culture yielded osteoblast-like cells which induced the formation of a higher number of osteoclast-like cells with a larger area compared to the explant culture with enzymatic pre-treatment when cultured with exogenous M-CSF and RANKL.
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2

Cramer, E. E. A., K. Ito, and S. Hofmann. "Ex vivo Bone Models and Their Potential in Preclinical Evaluation." Current Osteoporosis Reports 19, no. 1 (January 11, 2021): 75–87. http://dx.doi.org/10.1007/s11914-020-00649-5.

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Abstract Purpose of Review Novel therapies for damaged and diseased bone are being developed in a preclinical testing process consisting of in vitro cell experiments followed by in vivo animal studies. The in vitro results are often not representative of the results observed in vivo. This could be caused by the complexity of the natural bone environment that is missing in vitro. Ex vivo bone explant cultures provide a model in which cells are preserved in their native three-dimensional environment. Herein, it is aimed to review the current status of bone explant culture models in relation to their potential in complementing the preclinical evaluation process with specific attention paid to the incorporation of mechanical loading within ex vivo culture systems. Recent Findings Bone explant cultures are often performed with physiologically less relevant bone, immature bone, and explants derived from rodents, which complicates translatability into clinical practice. Mature bone explants encounter difficulties with maintaining viability, especially in static culture. The integration of mechanical stimuli was able to extend the lifespan of explants and to induce new bone formation. Summary Bone explant cultures provide unique platforms for bone research and mechanical loading was demonstrated to be an important component in achieving osteogenesis ex vivo. However, more research is needed to establish a representative, reliable, and reproducible bone explant culture system that includes both components of bone remodeling, i.e., formation and resorption, in order to bridge the gap between in vitro and in vivo research in preclinical testing.
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3

Wongwicha, Winida, Hiroyuki Tanaka, Yukihiro Shoyama, Indree Tuvshintogtokh, and Waraporn Putalun. "Production of Glycyrrhizin in Callus Cultures of Licorice." Zeitschrift für Naturforschung C 63, no. 5-6 (June 1, 2008): 413–17. http://dx.doi.org/10.1515/znc-2008-5-617.

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Licorice plants, Glycyrrhiza glabra, G. uralensis, and G. inflata, were investigated for callus induction using Murashige and Skoog (MS) medium combined with auxins and cytokinins. After 4 weeks of culture, 33-100% of leaf or stem explants formed calli. Maximum of shoot induction from callus cultures was achieved by G. inflata stem explants cultured on MS medium supplemented with 1 mg/l α-naphthaleneacetic acid (NAA) and 0.5 mg/l 6-benzyladenine (BA) (67%) which also gave maximum shoot formation per explant (two shoots per explant). These results indicated that all three Glycyrrhiza species regenerated shoots from callus cultures on MS medium combined with NAA and BA or only thidiazuron (TDZ; 0.1 and 0.5 mg/l). Glycyrrhizin contents of G. uralensis calli induced using MS medium in combination with NAA and BA [(27.60 ± 8.47) μg/g DW] or TDZ alone [(36.52 ± 2.45) μg/ g DW] were higher than those found in other combinations.
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4

Kahia, Jane, Margaret Kirika, Hudson Lubabali, and Sinclair Mantell. "High-frequency Direct Somatic Embryogenesis and Plantlet Regeneration from Leaves Derived from In Vitro-Germinated Seedlings of a Coffea arabica Hybrid Cultivar." HortScience 51, no. 9 (September 2016): 1148–52. http://dx.doi.org/10.21273/hortsci10771-16.

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Breeding work carried out during the period 1971–85 by the Coffee Research Institute, Ruiru, Kenya resulted in the release of a new improved hybrid Coffea arabica named Ruiru 11. The cultivar combines resistance to coffee berry disease (CBD) and leaf rust, with high yield and good cup quality attributes. The propagation by F1 hybrid seeds production, cuttings, and tip grafting do not produce enough planting materials. There was a need to explore alternative methods and tissue culture offers potential options. The objective of the study was to evaluate the effect of explant sources and cytokinins on induction and regeneration of somatic embryos. Eight different explants were cultured on half-strength Murashige and Skoog (MS) medium supplemented with 10 µm benzylaminopurine (BAP). The effect of kinetin, N6-(2-isopentyl) adenine (2iP) evaluated at (0, 0.5, 5, or 25 µm) or thidiazuron (TDZ) (0, 0.5, 1.0, or 5 µm) added in separate experiments was also evaluated. The percentage of embryogenic cultures and the numbers of embryos per explant were determined after 3 months’ culture. The explant type had a significant effect (P > 0.05) on the induction of somatic embryos. Explants from in vitro-germinated seedlings produced the highest embryogenic cultures (90%) and the highest mean number of embryos (19.36) per explant. Cytokinins strongly enhanced induction and regeneration of somatic embryos. TDZ at 1 µm produced the highest embryogenic cultures (100%) and the highest mean number of embryos (24.2). The embryos were germinated on half-strength MS medium without any hormones. A high (98%) survival rate of the regenerated plantlets was recorded over all the treatments in the greenhouse. This is the first report on induction of high-frequency direct somatic embryos from coffee juvenile tissues. This is of great significance in tissue culture and indeed molecular biology manipulations because it allows regeneration of coffee from several explants.
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5

Chowdhary, Ashish K., and S. N. Bahuguna. "Development of primary cell culture from the caudal fin region of snow trout Schizothorax plagiostomus (Heckel)." Environment Conservation Journal 15, no. 1&2 (June 18, 2014): 149–52. http://dx.doi.org/10.36953/ecj.2014.151219.

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A primary cell culture has been established from the Snow trout Schizothorax plagiostomus. Primary cultures have developed from caudal fin tissues by explant culture technique. Cell outgrowth has been obtained from the caudal fin explant after 5-7 days of explant culture. The culture medium used for the experiment was Leibovitz-15 supplemented with 20% Fetal Bovine Serum. Radiation of cells from the explants started after 2 to 3 days. Both fibroblast like and epithelial like cells were noticed but fibroblast like cells dominated as the growth progressed.
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6

Plessis, Helena Jacoba du, Roumiana Vassileva Nikolova, Bronwyn Anne Egan, and Riana Kleynhans. "Preliminary study on in vitro shoot culture of Hibiscus coddii subsp. barnardii, an indigenous South African flowering plant." Ornamental Horticulture 27, no. 3 (September 2021): 408–16. http://dx.doi.org/10.1590/2447-536x.v27i3.2353.

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Abstract In vivo and in vitro grown plants of Hibiscus coddii subsp. barnardii were used as explant source for establishment of in vitro cultures. Nodal shoot explants derived from in vivo grown plants, both naturally and under controlled environmental conditions, showed high sensitivity to the surface disinfection treatment and poor survival in in vitro culture. In vitro grown seedlings proved successful as aseptic source of apical and basal shoot explants to establish contamination-free in vitro cultures. Sprouting of axillary buds was observed on 90% of apical shoot explants after four weeks of culture on full strength, plant growth regulator (PGR)-free Murashige and Skoog (MS) medium. However, further proliferation of short shoots, limited to the bud sprout at the explant base, occurred on only 50% of these explants. In contrast, all basal shoot explants attained 3-5 single primary axillary shoots (30-40 mm in length) while a clump of short (5-10 mm) shoots also formed at the base in 60% of these explants. In both explant types, addition of 0.25-1 mg L-1 6-Benzylaminopurine (BAP) to the MS medium resulted in a low frequency (10%-60%) of explants with short shoots (5-10 mm) that showed no further elongation. Moreover, explants cultured in the presence of BAP showed a high frequency of callus formation (up to 90%) and low survival (20%-60%). A lower frequency of callus formation (30%-40%) and higher survival (90%-100%) of both explant types occurred on BAP-free medium. Further subculturing of primary and secondary axillary shoots onto fresh MS medium (with and without BAP) did not improve shoot multiplication. Regenerated plantlets from PGR-free MS medium were successfully acclimatized and hardened-off.
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7

Heger, Julia I., Karolin Fröhlich, Lisa Uhl, Andre Schmidt, and Udo R. Markert. "Toxicological analyses on placental explant cultures." Placenta 57 (September 2017): 254. http://dx.doi.org/10.1016/j.placenta.2017.07.108.

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8

Cao, X., and F. Hammerschlag. "307 Growth Regulator Pretreatments Significantly Enhance the Efficiency of Shoot Organogenesis from Leaf Explants of Highbush Blueberry Cultivar Bluecrop." HortScience 35, no. 3 (June 2000): 445A—445. http://dx.doi.org/10.21273/hortsci.35.3.445a.

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As part of a program to develop transgenic highbush blueberry (Vaccinium corymbosum L.) cultivars, studies were conducted to determine optimum conditions for high-efficiency shoot regeneration from leaf explants of in vitro propagated, commercially important, tissue culture-recalcitrant `Bluecrop' shoot cultures. The effects of pretreatments, growth regulators, and age of explant source on shoot organogenesis were investigated. A maximum of 98% shoot regeneration and 10 shoots regenerating per leaf explant occurred when explants of 2-week-old shoot cultures were incubated in the dark (for a total of 14 days) on pretreatment medium #1 containing 2.6 μM NAA and 5 μM TDZ for 4 days, next on pretreatment medium #2 containing 2.6 μM NAA and 7 μM zeatin riboside for 3 days, then on regeneration medium containing 1 μM TDZ for 6 weeks, and last on medium without growth regulators for 10 days. No shoot regeneration occurred if explants were incubated without exposure to pretreatments before incubation on regeneration medium. There were no significant differences in percentage of regeneration or the number of shoots regenerating per explant from leaf explants derived from either 1-, 2-, or 3-week-old shoot cultures. Shoot production per explant on 1 μM TDZ was about three times that on either 0.5 μM TDZ or 20 μM zeatin riboside, and nine times that on 5 μM TDZ.
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9

Husna, Miftahul, Dimas Ramadhian Noor, Rizki Sekar Arum, Hana Qanita, Abinawanto, Anom Bowolaksono, Erwin Danil Julian, and Astari Dwiranti. "The Use of Patient-Derived Explant Cultures for Predicting Breast Cancer Cell Migration Potential In Vitro." Journal of Hunan University Natural Sciences 49, no. 5 (May 30, 2022): 145–50. http://dx.doi.org/10.55463/issn.1674-2974.49.5.16.

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Breast cancer ranks as one of the leading causes of death globally, while mortality rates due to breast cancer continue to rise. In Indonesia, breast cancer is the most common type of cancer, with women dying from breast cancer at a higher rate than men. Primary cultures play a significant role in examining the behavior of breast cancer cells. Because explant cultures can be performed directly from the primary tumor, they constitute a promising new tool for observing cell migration in cancer, including breast cancer. Nevertheless, the use of explant cultures to predict the migration of breast cancer cells has yet to be investigated. This study aims to assess the potential of the explant culture method to predict the migration ability of breast cancer cells in vitro. Tumor explants from two different patients were evaluated in this study. The explant cultures were observed for 14 days until passage, and the results were examined using a microscope. We found that BC02 cells took less than seven days to migrate from the primary tumor, while BC01 cells took 21 days. Furthermore, a mammosphere was observed in the BC02 sample. The rate of cell migration from the tissue depends on the malignant status of the tissue. In conclusion, this study suggests that explant cultures can be used to study the characteristics of cancer cell migration and its correlation with the malignancy of the original tissue.
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10

TANG, CHU-NIE, DHILIA UDIE LAMASUDIN, WAN MUHAMAD ASRUL NIZAM WAN ABDULLAH, CHEW-LI MOO, MIAO-SI CHIEW, QIAN-YEE CHAI, JANNA ONG-ABDULLAH, and KOK-SONG LAI. "Enhanced in vitro Shoot Regeneration and Biochemical Properties of Stevia rebaudiana using Chitosan." Sains Malaysiana 50, no. 3 (March 31, 2021): 667–76. http://dx.doi.org/10.17576/jsm-2021-5003-09.

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Stevia rebaudiana is a herbaceous perennial plant with great global demand due to its beneficial steviol glycosides(SGs) content. Current conventional breeding technique is unable to cater the need for more S. rebaudiana planting materials. Therefore, an improved in vitro shoot regeneration protocol was developed for S. rebaudiana by using chitosan. The highest fresh weight of plant (0.586 ± 0.176 g/explant), dry weight of plant (0.056 ± 0.02 g/explant) and plant height (4.94 ± 1.17 cm/explant) with maximum number of leaves (25.33 ± 6.95 /explant) were observed on explants grown in optimun treatment of MS basal medium supplemented with 1.0 mg/L of 6-benzyaminopurine (BAP) and 60 mg/L of low molecular weight (MW) chitosan after 4 weeks of culture. Scanning electron microscopy (SEM) analysis showed that new shoot primordia and shoot bud formation can be seen as early as day 3 and 5 of cultures on optimun treatment. Further biochemical assays showed that total phenolic acid content, total protein content and total hydrolyzed sugar content were recorded higher in explants cultured in optimun treatment as compared to control media. In contrast, total chlorophyll content and total flavonoids content were reduced in optimum treatment. Meanwhile, no significant difference in antioxidant activity was observed. All cultures from the optimal treatment were successfully regenerated, acclimatized and grew well with 100% survival rate. Taken together, an enhanced and efficient shoot regeneration protocol of S. rebaudiana was successfully developed which will be useful for rapid and large-scale micropropagation in future.
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11

Arum, Rizki Sekar, Dimas Ramadhian Noor, Miftahul Husna, Hana Qanita, Abinawanto, Anom Bowolaksono, Erwin Danil Julian, and Astari Dwiranti. "Deriving Breast Cancer’s Primary Cultures from Patients' Tumor Biopsies in Indonesia Using Explant and Enzymatic Methods." Journal of Hunan University Natural Sciences 49, no. 9 (September 30, 2022): 11–16. http://dx.doi.org/10.55463/issn.1674-2974.49.9.2.

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Breast cancer is the most common cause of death in women globally with high rates of heterogeneity. The development of breast cancer treatment is still constrained because of the different responses to its therapy. The use of primary breast cancer culture is invaluable because it provides the same properties as breast cancer itself. Primary culture is used as a tool to determine the proliferative ability of breast cancer cells. However, the use of optimum methods in cultivating primary culture must be evaluated because of the unstable nature of primary culture. In this study, we investigate the optimum method for culturing cells derived from the tumor biopsies of breast cancer patients in Indonesia by comparing explant and enzymatic methods. Breast cancer tissues were obtained from five breast cancer patients, who underwent surgery and incision biopsies. Tissues were further cultured by explant and enzymatic methods. The cell cultures were observed daily using a microscope for up to 30 days. The results showed that the cells cultured using the enzymatic method for more than 16 h were susceptible to microbiome contamination in the following days after enzymatic digestion, while those cultured using the explant method grew well for 30 days. The findings of this research suggested that the explant method gave better results compared with the enzymatic method. The findings of this study provide insight into the optimum conditions for the primary culture of breast cancer cells.
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Rybczyński, Jan J., Marta Karolkowska, Zygmunt Kaczmarek, Anna Mikuła, and Agnieszka Fiuk. "In vitro morphogenic events in culture of Lotus corniculatus L. seedling root explants." Acta Societatis Botanicorum Poloniae 75, no. 3 (2011): 191–200. http://dx.doi.org/10.5586/asbp.2006.022.

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The experiments were carried out on <em>Lotus corniculatus</em> (L.) seedling root explants of the cultivar varieties Skrzeszowicka, Caroll A10 and strain 175. Callus formation and shoot regeneration were the major explant response depended mainly on of the studied genotype and used plant growth regulators (PGRs). Primary cortex of proximal and distal end of explant was the most active tissue for callus proliferation. For shoot primordia differentiation deeper zones of cortex took a part. The process of meristematic centre initiation was not uniform and various level of shoot differentiation events were observed not earlier than 3 weeks of culture. Usually, the shoot primordia regeneration began on proximal rather than distal end of the explant. BAP rather than urea derivatives stimulated shoot proliferation in extended cultures. Increasing of BAP and TDZ concentrations brought about the explant polarity and expansion of the meristematic zones. The explant position in root did not have significant influence on the number of regenerated shoots. The cultures only had better bud formation by TDZ when compared to BAP. BAP stimulated bud formation and development of the shoots from them. Short term of TDZ treatment of explants stimulated meristem formation which developed into buds and shoots. CPPU stimulated callus proliferation and bud formation when explants pretreatment was prolonged from 12 to 36 hrs.
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13

Gutnick, M. J., B. Wolfson, and F. Baldino. "Synchronized neuronal activities in neocortical explant cultures." Experimental Brain Research 76, no. 1 (June 1989): 131–40. http://dx.doi.org/10.1007/bf00253630.

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14

Martini, Aekaterini N., Maria Papafotiou, and Stavros N. Vemmos. "Season and Explant Origin Affect Phenolic Content, Browning of Explants, and Micropropagation of ×Malosorbus florentina (Zucc.) Browicz." HortScience 48, no. 1 (January 2013): 102–7. http://dx.doi.org/10.21273/hortsci.48.1.102.

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The aim of this study was to develop an efficient protocol for in vitro propagation of the rare and endangered ×Malosorbus florentina, not only enabling conservation of the species, but also its use as an ornamental. Explants excised from adult plants, shoot tip explants, and explants collected in March and April showed more browning and had higher content of total phenolics than explants excised from juvenile tissue, nodal explants, and those collected during any of the other months of the year. Shoot tip explants from adult plants were more difficult to establish in vitro (14%) compared with explants from micropropagated plantlets or sprouts of burned plants (29% to 36%). Nodal explants excised from seedlings were established at the highest percentage (83%), giving the most shoots per explant (5.2). Generally, in vitro cultures established from adult plants, with the exception of one culture, showed lower multiplication rates compared with cultures from juvenile plants. Nodal explants from the base of sprouts produced a higher percentage (60%) of shoots than explants from upper locations (20% to 31%), but any differences in proliferation rates of established cultures ceased after the third subculture. Microshoots from juvenile cultures were more capable of rooting (51% to 58%) than were those from adult plants (16% to 32%), whereas 83% of the plantlets were acclimatized ex vitro independently of their origin, but plantlets of juvenile origin, although developing the same height as those originating from adult plants, had shorter internodes and thus more compact shape.
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Mohamed, Mohamed F., Paul E. Read, and Dermot P. Coyne. "Dark Preconditioning, CPPU, and Thidiazuron Promote Shoot Organogenesis on Seedling Node Explants of Common and Faba Beans." Journal of the American Society for Horticultural Science 117, no. 4 (July 1992): 668–72. http://dx.doi.org/10.21273/jashs.117.4.668.

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Dry seeds from two lines of common bean (Phaseolus vulgaris L.) and one cultivar of faba bean (Vicia faba L.) were germinated on Murashige and Skoog (MS) medium containing B vitamins, 30 g sucrose/liter, and either 2.5, 5.0, or 7.5 μm benzyladenine (BA). Axenic seed cultures were grown at 22 to 24C in darkness and under continuous light from cool-white fluorescent tubes (40 μmol·m-2·s-1). Explant tissues were prepared from cotyledonary nodes (CN) and primary nodes (PN) of 14-day-old seedlings. Explants were cultured on corresponding seedling growth medium and maintained under continuous cool-white light (40 μmol·m-2·s-1). The percentages of CN and PN (in one line of common bean) explants that regenerated shoots and the number of shoots per explant (in all germplasm) were highest when nodal tissues were prepared from seedlings germinated in darkness. These responses were optimal on medium containing 5 μm BA during seedling growth and subsequent culture of explants. The number of shoots per explant was two to five times higher on explants cultured on medium with 0.25 to 1.0 μm forchlorfenuron (CPPU) or thidiazuron (TDZ) than on medium with 5 μm BA. Higher (2.5 and 5 μm) CPPU and TDZ concentrations inhibited shoot elongation and stimulated callus production. Histological analyses indicated that adventitious meristems formed 6 to 8 days after explant culture. Progenies from regenerated plants appeared similar to plants raised from the original seed stocks. Chemical names used: N- (phenylmethyl) -1 H- purin-6-amine (benzyladenine, BA); N- (2-chloro-4-pyridyl)-N'- phenylurea (forchlorfenuron, CPPU); N- phenyl -N' -1,2,3-thiadiazol-5-ylurea (thidiazuron, TDZ).
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Vila, Inmaculada, Ester Sales, Javier Ollero, Jesús Muñoz-Bertomeu, Juan Segura, and Isabel Arrillaga. "Micropropagation of Oleander (Nerium oleander L.)." HortScience 45, no. 1 (January 2010): 98–102. http://dx.doi.org/10.21273/hortsci.45.1.98.

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Successful propagation of Nerium oleander L. (oleander) was achieved by in vitro methods. Shoot cultures were initiated from seedlings of wild-growing plants and from shoot apices of adult plants belonging to the commercial cultivars Splendens Giganteum, Revanche, and Alsace. Axillary shoot breaking from shoot tips excised from these cultures required the presence of either 6-benzylaminopurine (BA) or thidiazuron (TDZ). The higher number of axillary shoots from juvenile material was obtained by culturing shoot tips from BA-pretreated material derived from seedlings on a modified Schenk and Hildebrandt medium (SHM) supplemented with BA or TDZ (average of 3.9 shoots per explant with a mean length of 10.4 mm) and when the media were supplemented with 8.8 μM TDZ (average of 3.5 shoots per explant with a mean length of 7.3 mm) or 4.4 μM BA (average of 3.3 shoots per explant with a mean length of 12.3 mm). Among cultivars, cv. Revanche showed best shoot proliferation rates, especially when explants were cultured on Woody Plant Medium (average of 3.2 shoots per explant with a mean length of 10.2 mm). Adventitious bud differentiation from oleander leaves is also reported. Leaves excised from seedling-derived shoot cultures responded better than those from adult plant-derived shoot cultures (40% versus 5%, respectively). Bud differentiation required the presence of TDZ in the SHM medium, although shoot development was only achieved on transference of explants to media without TDZ but supplemented with BA and indoleacetic acid (IAA) or BA, kinetin, and IAA. Axillary and adventitious shoots were easily rooted (99%) and successfully (95% to 100%) transferred to soil.
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Cao, Xiaoling, Freddi A. Hammerschlag, and Larry Douglass. "A Two-step Pretreatment Significantly Enhances Shoot Organogenesis from Leaf Explants of Highbush Blueberry cv. Bluecrop." HortScience 37, no. 5 (August 2002): 819–21. http://dx.doi.org/10.21273/hortsci.37.5.819.

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As part of a program to improve highbush blueberry (Vaccinium corymbosum L.) cultivars via tissue culture and genetic engineering, studies were conducted to determine optimum conditions for organogenesis from leaf explants of the previously recalcitrant cv. Bluecrop. The effects of a pretreatment, growth regulators, and age of explant source on shoot organogenesis were investigated. A maximum of 98% explants regenerated shoots with a mean of 11 shoots per leaf explant after 62 days when explants of 2-week-old shoot cultures were incubated on the following regime: pretreatment medium #1 containing 5 μm TDZ and 2.6 μm NAA for 4 days, pretreatment medium #2 containing 7 μm zeatin riboside and 2.6 μm NAA for 3 days, regeneration medium containing 1 μm TDZ for 6 weeks, and last on medium without growth regulators for 10 days. No shoot regeneration occurred if explants were incubated without exposure to pretreatment prior to incubation on regeneration medium. There were no significant differences in percentage of regeneration or the number of shoots regenerating per explant from leaf explants derived from either 1-, 2-, or 3-week-old shoot cultures. Shoot production per explant on regeneration medium containing 1 μm TDZ was about three times that on 0.5 μm TDZ or 20 μm zeatin riboside, and nine times that on 5 μm TDZ. Chemical names used: 1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea (thidiazuron, TDZ); 9-(β-D-ribofuranosyl)-6-(4-hydroxy-3-methyl-but-2-enylamino)purine (zeatin riboside).
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Kurek, Magdalena, Elisabet Åkesson, Masahito Yoshihara, Elizabeth Oliver, Yanhua Cui, Martin Becker, João Pedro Alves-Lopes, et al. "Spermatogonia Loss Correlates with LAMA 1 Expression in Human Prepubertal Testes Stored for Fertility Preservation." Cells 10, no. 2 (January 27, 2021): 241. http://dx.doi.org/10.3390/cells10020241.

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Fertility preservation for male childhood cancer survivors not yet capable of producing mature spermatozoa, relies on experimental approaches such as testicular explant culture. Although the first steps in somatic maturation can be observed in human testicular explant cultures, germ cell depletion is a common obstacle. Hence, understanding the spermatogonial stem cell (SSC) niche environment and in particular, specific components such as the seminiferous basement membrane (BM) will allow progression of testicular explant cultures. Here, we revealed that the seminiferous BM is established from 6 weeks post conception with the expression of laminin alpha 1 (LAMA 1) and type IV collagen, which persist as key components throughout development. With prepubertal testicular explant culture we found that seminiferous LAMA 1 expression is disrupted and depleted with culture time correlating with germ cell loss. These findings highlight the importance of LAMA 1 for the human SSC niche and its sensitivity to culture conditions.
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Vinterhalter, D., and Branka Vinterhalter. "Tropic responses of potato single-node explant cultures." Archives of Biological Sciences 64, no. 1 (2012): 183–89. http://dx.doi.org/10.2298/abs1201183v.

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A special in-vitro protocol was elaborated enabling the production of potato single-node explant plantlets that can be used as objects for tropic studies. In light-grown plantlets, achievement of a full (90?) phototropic (PT) curvature required 75 to 120 min of continuous unilateral blue light irradiation or 120-135 min of gravitropic stimulation (GT). Time-lapse photography revealed that the curves describing PT and GT bending have a sigmoid shape. Continuous BL irradiation was necessary for the induction of continuous PT bending. If the BL was turned off after 30-50 min of PT stimulation, the bending gradually decreased and stopped in darkness after 25.0 ? 2.0 min. Within this period, curvature increased by 15.5 ? 1.5?. When the BL was turned off upon completion of PT bending (when the plantlets reached an angle of 90?), the plantlets entered the phase of fast straightening. The 90? PT curvature was significantly exaggerated in darkness by turning the jars from a vertical to horizontal position providing 120.74 ? 2.5? as the final curvature angle after two more hours in darkness.
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Ironside, James W., Robert D. E. Battersby, John Lawry, Reginald S. Loomes, Christopher A. Day, and Walter R. Timperley. "DNA in meningioma tissues and explant cell cultures." Journal of Neurosurgery 66, no. 4 (April 1987): 588–94. http://dx.doi.org/10.3171/jns.1987.66.4.0588.

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✓ Flow cytometry was performed on stored frozen tissues and explant cell cultures from 39 meningiomas using ethidium bromide and mithramycin in a selective staining technique for deoxyribonucleic acid (DNA). The ploidy index and percentage of cells in the G0/G1, S, and G2/M phases were calculated for each specimen. The results were compared with the age and sex of the patients; the site, the histological subtype, and mitotic rate of the neoplasms; and the estrogen and progesterone-receptor levels assayed in cytosol-enriched supernatants from cryostat-cut sections. Sixteen neoplasms (41%) were aneuploid. These included two recurrent neoplasms, seven of the eight neoplasms from patients with multiple meningiomas, and three clinically aggressive neoplasms (one hemangiopericytic and two anaplastic meningiomas). Significant correlations were found between values for the ploidy index (r =0.75, p < 0.01), the percentage of S-phase cells (r = 0.82, p < 0.01), and the percentage of G2/M-phase cells (r = 0.69, p < 0.05) in vivo and in vitro. The results support the suggestion that flow cytometry for DNA in meningiomas may be of value in predicting the behavior of these neoplasms, and indicate that under controlled conditions explant cell cultures may provide a useful model for the proliferative characteristics of meningiomas in vivo.
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Orth, M. W., T. L. Peters, and K. A. Chlebek-Brown. "Cartilage Turnover in Embryonic Chick Tibial Explant Cultures." Poultry Science 79, no. 7 (July 2000): 990–93. http://dx.doi.org/10.1093/ps/79.7.990.

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22

DePasquale, Joseph A. "Rodlet cells in epidermal explant cultures ofLepomis Macrochirus." Acta Zoologica 95, no. 2 (November 28, 2012): 144–54. http://dx.doi.org/10.1111/azo.12013.

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23

Baumgartner, J. Craig, and William A. Falkler. "Biosynthesis of IgG in periapical lesion explant cultures." Journal of Endodontics 17, no. 4 (April 1991): 143–46. http://dx.doi.org/10.1016/s0099-2399(06)82005-4.

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24

Neubauer, J. A., S. F. Gonsalves, W. Chou, H. M. Geller, and N. H. Edelman. "Chemosensitivity of medullary neurons in explant tissue cultures." Neuroscience 45, no. 3 (January 1991): 701–8. http://dx.doi.org/10.1016/0306-4522(91)90282-s.

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25

Behera, Biswaranjan, Shashikanta Behera, Padan K. Jena, Durga P. Barik, and Soumendra K. Naik. "Adventitious Shoot Organogenesis and Plant Regeneration from Internode Explants of Paederia foetida L.:A Valuable Medicinal Plant." Biosciences, Biotechnology Research Asia 14, no. 3 (September 25, 2017): 893–900. http://dx.doi.org/10.13005/bbra/2523.

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ABSTRACT: A plant regeneration protocol via adventitious shoot organogenesis from internode explants of Paederia foetida (Skunk vine) is reported here for the first time. Three explants (leaf, mature internode and internode derived from axenic shoot cultures) were tested for shoot organogenesis. Leaf explants failed to induce adventitious shoots whereas axenic internode explant was found to be superior to mature internode explants for the induction of adventitious shoots. Axenic internode explants cultured on MS medium supplemented with 3.0 mg/l BAP showed maximum (86.7 %; 10.4 shoots per explant) adventitious shoot organogenesis. The regenerated shoots were best rooted (90 %; 14 roots per shoot) on half-strength MS medium. Eighty percent of the rooted shoots were successfully acclimatized in soil: sand (1:1) mixture. All these acclimatized plants were successfully transferred to larger pots containing garden soil and subsequently established in the field.
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Dewir, Yaser Hassan, Abdulla Alsadon, Ahmed Ali Al-Aizari, and Mohaidib Al-Mohidib. "In Vitro Floral Emergence and Improved Formation of Saffron Daughter Corms." Horticulturae 8, no. 10 (October 20, 2022): 973. http://dx.doi.org/10.3390/horticulturae8100973.

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In vitro cormogenesis is a potential tool for improving saffron production under controlled conditions. In this study, the effects of explant type, culture type, and medium supplements on saffron daughter corm formation in vitro were assessed. Saffron flowers emerged 30 days after culture, and the sizes of in-vitro- and ex-vitro-produced flowers and stigmas were similar. In vitro daughter corm formation and the saffron life cycle was completed after 10 and 14 weeks of culture, respectively. Using in vitro intact corms was more effective for corm production than using apical buds. Compared with apical bud explants, mother corm explants produced more corms with a higher fresh weight and diameter. Compared with solid culture, liquid cultures using bioreactors provided corms with a higher fresh weight and diameter, regardless of explant type. An ebb and flow system provided the highest cormlet fresh weight and diameter but the fewest cormlets, whereas an immersion system provided more cormlets with a smaller size. Saffron apical buds cultured with salicylic acid at 75 mg L−1 or glutamine at 600 mg L−1 exhibited the highest cormlet diameter and fresh weight. These findings will improve the process of in vitro cormogenesis and the production of saffron under controlled conditions.
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Shekarian, Tala, Ewelina Bartoszek-Kandler, Carl Zinner, Christian Schuerch, and Gregor Hutter. "EXTH-40. MULTIDIMENSIONAL INTERROGATION OF GLIOBLASTOMA MICROENVIRONMENT TREATMENT RESPONSE IN EXPLANT CULTURES." Neuro-Oncology 23, Supplement_6 (November 2, 2021): vi172. http://dx.doi.org/10.1093/neuonc/noab196.679.

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Abstract The immune tumor microenvironment (iTME) of glioblastoma (GBM) contains microglial, macrophage, other myeloid cell populations and as adaptive immune cells. Recent therapeutic strategies for GBM aim at targeting iTME components to induce antitumoral immunity. A patient-tailored, ex vivo drug testing and response analysis platform would facilitate personalized therapy planning, provide insights into treatment-induced immune mechanisms in the iTME, and enable the discovery of biomarkers of response and resistance. Here, we generated patient-derived, live 3D GBM bioreactors from different tumor regions to assess iTME treatment responses to microglia modulators and immune checkpoint inhibitors. Intact GBM tissue specimens from the tumor center and periphery were cultured for 7 days in the presence or absence of anti-PD1, anti-CD47 antibodies or their combination. Tissues were analyzed by CODEX highly multiplexed microscopy using an immune-centered 54-marker panel, and changes in cytokine and chemokine levels in culture supernatants were investigated. A computational pipeline for integrative therapy response assessment was implemented. Explant cultures from n=8 IDH wt GBM were subjected to this integrative personalized analysis. Tissue integrity after 3D bioreactor cultures was comparable to tissue taken directly after surgery. FFPE CODEX workflow was feasible with adequate staining quality in bioreactor cultures. 850'000 single cells were segmented and clustered. Cellular composition between tumor center and the peripheral invasion zone differed significantly in immune phenotypes, cytokine profile and response to innate, adaptive or combinatorial local immunotherapies. Multiplexed cytokine analysis revealed IFNγ response signatures in a subset of center samples, whereas the peripheral invasion zone displayed a blunted cytokine response. This cytokine signature corresponded to cellular composition shifts within specific cellular neighborhoods. CD4 and CD8 T cells were invigorated and left their vascular niche. Our study demonstrates that local immunotherapies enable an active antitumoral immune response within the tumor center, and provides a multidimensional personalized framework for immunotherapy response assessment.
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Gaj, Małgorzata D., Grzegorz Czaja, and Małgorzata Nawrot. "Selection of valine-resistance in callus culture of Arabidopsis thaliana (L.) Heynh. derived from leaf explants." Acta Societatis Botanicorum Poloniae 68, no. 3 (2014): 211–15. http://dx.doi.org/10.5586/asbp.1999.028.

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The selection of valine-resistant mutants was carried out in leaf explant cultures of three <em>Arabidopsis thaliana</em> (L.) Heynh. ecotypes: C-24, RLD and Columbia. The valine concentration used for in vitro selection, lethal for seed-growing plants, has not affected callus formation and growth. However, strong inhibition of shoot regeneration ability of calli growing under selection pressure was noticed. In total, 1043 explants were cultured on valine medium and 18 shoots were regenerated with an average frequency of 1.7 shoots per 100 calli. Most R<sub>1</sub> shoots were sterile and seeds were collected from 3 plants. The transmission of valine-resistance to the sexual progeny of these plants was scored and the increased level of valine-resistance was found in progeny of one line - 61 C. This line originated from the culture of Columbia leaf explant and displayed tetraploid chromosome number.
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Chen, L., S. O. Park, S. Dhir, and A. S. Bhagsari. "331 Effects of Explant Type, Sucrose Level and Callus Development Time on In Vitro Plant Regeneration of Sweetpotato." HortScience 35, no. 3 (June 2000): 449A—449. http://dx.doi.org/10.21273/hortsci.35.3.449a.

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Several experiments were conducted to evaluate the influence of explant type, sucrose level, and callus development time on sweetpotato [Ipomoea batatas (L.) Lam] in vitro culture. Shoot tip, petiole, and leaf of Selection 75-96-1 was used as explants in Murashige and Skoog (MS) media with different plant growth regulators. Calli derived from shoot tip and petiole produced 42.1% and 10.3% somatic embryos, respectively, but the leaf failed to produce somatic embryos. The effect of sucrose level was determined using shoot tip as explants. Compared with 3% sucrose in the same plant growth regulators level medium during callus initiation and callus proliferation periods, 5% sucrose level suppressed root growth and improved shoot regeneration. The callus development time was measured by using shoot tips on callus initiation medium containing 1.5 mg/L alpha-Naphthaleneacetic acid (NAA) and 0.25 mg/L Kinetin (KIN) plus 5% sucrose. When explants were cultured for less than 6 weeks during callus initiation, then transferred onto plant regeneration medium, plant regeneration via organogenesis occurred; whereas, maintaining cultures for more than 12 weeks on the same callus initiation/proliferation medium, plant regeneration was favored via embryogenesis. Explant type and other factors affecting plant regeneration noted here could be applied to protoplast culture, somatic hybridization, and transformation in sweetpotato.
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30

Seabrook, Janet E. A., and Gerald Farrell. "City Water Can Contaminate Tissue Culture Stock Plants." HortScience 28, no. 6 (June 1993): 628–29. http://dx.doi.org/10.21273/hortsci.28.6.628.

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Stock plants of `Shepody' and `Yukon Gold' potato (Solarium tuberosum L.) were grown in a greenhouse and irrigated with city water. Contamination rate of stem explant tissue cultures excised from these stock plants was 50% to 100%. A comparison of the microorganisms isolated from the contaminated cultures and from 0.22-μm filter disks through which 20 liters of city water had passed revealed the presence of similar bacterial floras. Five genera of bacteria (Listerium spp., Corynebacterium spp., Enterobacter spp., Pasteurella spp., and Actinobacillus spp.) were isolated from contaminated cultures and cultured filter disks. Watering greenhouse-grown stock plants with filtered city water decreased contamination of stem explant cultures 30% to 50%. Installing an ultraviolet light water-sterilizing unit at the greenhouse inlet point effectively reduced contamination.
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31

Dhekney, Sadanand A., Zhijian T. Li, Michael E. Compton, and Dennis J. Gray. "Optimizing Initiation and Maintenance of Vitis Embryogenic Cultures." HortScience 44, no. 5 (August 2009): 1400–1406. http://dx.doi.org/10.21273/hortsci.44.5.1400.

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Stamens and pistils from mature grapevines and leaves from in vitro micropropagation cultures were used to optimize parameters influencing somatic embryogenesis in Vitis. Embryogenic competence was dependent on species/variety, explant type and developmental stage, medium composition, and growth regulator concentration. Of varieties evaluated, a greater number produced embryogenic cultures from stamens and pistils (26) compared with leaves (six). Among the different stamen and pistil stages, Stage II and III explants produced the maximum embryogenic response regardless of genotype and medium composition. Of seven culture media tested, the highest embryogenic response was recorded from varieties cultured on MSI (18) and PIV (16) media. Experiments annually repeated over 3 to 10 years demonstrated reproducible results. Highly reliable protocols for somatic embryogenesis were obtained for 29 Vitis species and varieties, including 18 Vitis vinifera varieties, Vitis riparia, Vitis rupestris, Vitis champinii, and eight Vitis hybrids. Embryogenic cultures were maintained on X6 medium for a period of 6 months to 2 years depending on the variety and used in studies involving genetic transformation and transgenic plant regeneration.
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Chen, Chien-Chih, Rick Bates, and John Carlson. "Effect of environmental and cultural conditions on medium pH and explant growth performance of Douglas-fir (Pseudotsuga menziesii) shoot cultures." F1000Research 3 (May 8, 2015): 298. http://dx.doi.org/10.12688/f1000research.5919.2.

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The medium pH level of plant tissue cultures has been shown to be essential to many aspects of explant development and growth. Sensitivity or tolerance of medium pH changein vitrovaries according to specific requirements of individual species. The objectives of this study are to 1) determine medium pH change over time in storage conditions and with presence of explants, 2) evaluate the effects of medium pH change on explant growth performance and 3) assess the effects of adding a pH stabilizer, 2-(N-morpholino)ethanesulfonic acid (MES) that is commonly used in Douglas-fir micropropagation medium. Vegetative buds were collected in the spring before breaking dormancy from juvenile and mature donor trees for conducting these evaluations. Medium, with or without MES, was pre-adjusted to five pH levels before adding MES, agar and autoclaving. Medium pH changes and explant growth parameters were measured at eight different incubation times. Overall, MES provided a more stable medium pH, relative to starting pH values, under both light and dark storage conditions as well as with presence of explants. A general trend of decreasing medium pH over time was found comparing explants from juvenile and mature donor genotypes. Explant height and weight growth increased over time, but differ among explants from juvenile and mature donor genotypes. Our findings suggest that a 21-day subculture practice may best sustain medium freshness, medium pH level and desirable explant growth.
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Chato-Astrain, Jesús, David Sánchez-Porras, Óscar Darío García-García, Claudia Vairo, María Villar-Vidal, Silvia Villullas, Indalecio Sánchez-Montesinos, Fernando Campos, Ingrid Garzón, and Miguel Alaminos. "Improvement of Cell Culture Methods for the Successful Generation of Human Keratinocyte Primary Cell Cultures Using EGF-Loaded Nanostructured Lipid Carriers." Biomedicines 9, no. 11 (November 6, 2021): 1634. http://dx.doi.org/10.3390/biomedicines9111634.

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Human skin keratinocyte primary cultures can be established from skin biopsies with culture media containing epithelial growth factor (EGF). Although current methods are efficient, optimization is required to accelerate the procedure and obtain these cultures in less time. In the present study, we evaluated the effect of novel formulations based on EGF-loaded nanostructured lipid carriers (NLC). First, biosafety of NLC containing recombinant human EGF (NLC-rhEGF) was verified in immortalized skin keratinocytes and cornea epithelial cells, and in two epithelial cancer cell lines, by quantifying free DNA released to the culture medium. Then we established primary cell cultures of human skin keratinocytes with basal culture media (BM) and BM supplemented with NLC-rhEGF, liquid EGF (L-rhEGF), or NLC alone (NLC-blank). The results showed that cells isolated by enzymatic digestion and cultured with or without a feeder layer had a similar growth rate regardless of the medium used. However, the explant technique showed higher efficiency when NLC-rhEGF culture medium was used, compared to BM, L-rhEGF, or NLC-blank. Gene expression analysis showed that NLC-rhEGF was able to increase EGFR gene expression, along with that of other genes related to cytokeratins, cell–cell junctions, and keratinocyte maturation and differentiation. In summary, these results support the use of NLC-rhEGF to improve the efficiency of explant-based methods in the efficient generation of human keratinocyte primary cell cultures for tissue engineering use.
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Piovan, Anna, Raffaella Filippini, and Gabbriella Innocenti. "Coumarin Compounds in Coronilla scorpioides Callus Cultures." Natural Product Communications 9, no. 4 (April 2014): 1934578X1400900. http://dx.doi.org/10.1177/1934578x1400900415.

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Coronilla scorpioides (L.) W.D.J. Koch is known for producing several compounds with pharmaceutical interest, such as the hydroxycoumarins umbelliferone, scopoletin and daphnoretin, the dihydrofuranocoumarin marmesin, and the furocoumarin psoralen. In vitro callus cultures of C. scorpioides were established from hypocotyl, leaf, stem internode and root explants in order to evaluate the possibility of in vitro production of these active secondary metabolites. Calli were obtained with high frequency from all the explant types both in B5 and MS medium. However, after the third subculture, B5 medium, giving the best results, was selected for subsequent transfers. Homogeneous calli were kept either in darkness or in light. Chemical analyses showed that scopoletin and the intermediate products of the biogenetic pathway of psoralen, umbelliferone and marmesin, were always present in the calli and excreted into the media, while daphnoretin was never detected. Light seems to be a prerequisite for psoralen biosynthesis. Root-derived calli produced a significantly higher amount of psoralen (137.5 μg g−1 DW). Principal component analysis showed that umbelliferone, marmesin and psoralen contents are related to variables associated with different explant types.
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Souza, Fernanda Vidigal Duarte, Begoña Garcia-Sogo, Antonio da Silva Souza, Amparo Pérez San-Juán, and Vicente Moreno. "Morphogenetic response of cotyledon and leaf explants of melon (Cucumis melo L.) cv. Amarillo Oro." Brazilian Archives of Biology and Technology 49, no. 1 (January 2006): 21–27. http://dx.doi.org/10.1590/s1516-89132006000100003.

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Callus cultures from cotyledon and leaf explants of a Spanish cultivar of melon (Amarillo Oro) were tested for growth and morphogenic capacity on several culture media with different concentrations of IAA (indole-3-acetic acid) in combination with 1.0 mg.L-1 BA (6-benzylaminopurine) or 6.0 mg.L-1 KIN (kinetin). The best results were achieved with cotyledon explants. The leaf explants presented low bud formation capacity. Variability of organogenic response on cotyledons of different age (7, 5, 3 and 1-day-old) was evaluated. The age of explant had a significant influence on bud induction. Cotyledon explants from 7-day-old seedlings showed higher organogenic index and development of shoots when cultured onto MS medium supplemented with 1.5 mg.L-1 of IAA and 1.0 mg.L-1 of BA. The effect of cut type of cotyledonary explants on organogenic response was also investigated. Explants cut transversally showed the best results. The addition of copper sulfate in the culture medium promoted a qualitative improvement of the regenerated shoots.
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36

Ang, S. L., and J. Rossant. "Anterior mesendoderm induces mouse Engrailed genes in explant cultures." Development 118, no. 1 (May 1, 1993): 139–49. http://dx.doi.org/10.1242/dev.118.1.139.

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We have developed germ layer explant culture assays to study the role of mesoderm in anterior-posterior (A-P) patterning of the mouse neural plate. Using isolated explants of ectodermal tissue alone, we have demonstrated that the expression of Engrailed-1 (En-1) and En-2 genes in ectoderm is independent of mesoderm by the mid- to late streak stage, at least 12 hours before their onset of expression in the neural tube in vivo at the early somite stage. In recombination explants, anterior mesendoderm from headfold stage embryos induces the expression of En-1 and En-2 in pre- to early streak ectoderm and in posterior ectoderm from headfold stage embryos. In contrast, posterior mesendoderm from embryos of the same stage does not induce En genes in pre- to early streak ectoderm but is able to induce expression of a general neural marker, neurofilament 160 × 10(3) M(r). These results provide the first direct evidence for a role of mesendoderm in induction and regionalization of neural tissue in mouse.
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37

Rani, V. K. Jaimsha, P. V. Fijesh, and Jose Padikkala. "Micropropagation of Ophiorrhiza eriantha Wight. through Leaf Explant Cultures." Plant Tissue Culture and Biotechnology 20, no. 1 (August 30, 2010): 13–20. http://dx.doi.org/10.3329/ptcb.v20i1.5960.

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Leaf explants of Ophiorrhiza eriantha cultured in MS supplemented with the combination of NAA 4 mg/l and BA 0.5 mg/l induced higher callus growth. The maximum number of shoots were produced from callus on the MS supplemented with BA 5 mg/l. The regenerated shoots were transferred into the auxin containing medium for rooting and IBA 3 mg/l supplemented medium produced maximum number of roots per shoot. Camptothecin (anticancer drug) was isolated from O. eriantha wild grown plant and in vitro regenerated plants, and was confirmed by LC-MS-MS. The camptothecin content in wild grown plant, callus and regenerated plants were quantified by HPLC system. Key words: Camptothecin, Leaf explants, Ophiorrhiza eriantha, Micropropagation D.O.I. 10.3329/ptcb.v20i1.5960 Plant Tissue Cult. & Biotech. 20(1): 13-20, 2010 (June)
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38

Austin, Douglas J., Romana A. Nowak, and Elizabeth A. Stewart. "Onapristone Suppresses Prolactin Production in Explant Cultures of Leiomyoma." Gynecologic and Obstetric Investigation 47, no. 4 (1999): 268–71. http://dx.doi.org/10.1159/000010120.

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39

TELANG, NITIN T., DEBORAH M. AXELROD, H. LEON BRADLOW, and MICHAEL P. OSBORNE. "Metabolic Biotransformation of Estradiol in Human Mammary Explant Cultures." Annals of the New York Academy of Sciences 586, no. 1 Biochemistry (May 1990): 70–78. http://dx.doi.org/10.1111/j.1749-6632.1990.tb17791.x.

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40

TELANG, NITIN T., AMARESH BASU, MUKUND J. MODAK, and MICHAEL P. OSBORNE. "Cellular ras Protooncogene Expression in Human Mammary Explant Cultures." Annals of the New York Academy of Sciences 586, no. 1 Biochemistry (May 1990): 230–37. http://dx.doi.org/10.1111/j.1749-6632.1990.tb17811.x.

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41

Yamamoto, Rie, Hiroshi Katsuki, Toshiaki Kume, Shuji Kaneko, Akinori Akaike, Shinichi Manabe, Satoshi Kashii, and Yoshihito Honda. "Evaluation of retinal ganglion cell death using explant cultures." Japanese Journal of Pharmacology 82 (2000): 171. http://dx.doi.org/10.1016/s0021-5198(19)48146-8.

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42

BAUMGARTNER, J., and W. FALKLERJR. "Detection of immunoglobulins from explant cultures of periapical lesions." Journal of Endodontics 17, no. 3 (March 1991): 105–10. http://dx.doi.org/10.1016/s0099-2399(06)81739-5.

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43

Fröhlich, Karolin, Julia Heger, Andre Schmidt, Yvonne Heimann, Boodor Al-Kawlani, Amelie Lupp, Gitta Turowski, and Udo R. Markert. "Effects of therapeutic drugs on human placenta explant cultures." Placenta 45 (September 2016): 104. http://dx.doi.org/10.1016/j.placenta.2016.06.151.

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44

Sladek, Celia D., and Mark J. Gallagher. "Vasopressin gene expression in explant and dispersed hypothalamic cultures." Regulatory Peptides 45, no. 1-2 (April 1993): 47–52. http://dx.doi.org/10.1016/0167-0115(93)90181-7.

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45

Hendelman, W. J., N. Savigny, and K. C. Marshall. "Growth and myelination of explant cultures in defined medium." In Vitro Cellular & Developmental Biology 21, no. 2 (February 1985): 129–34. http://dx.doi.org/10.1007/bf02620955.

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46

Goncalves, Juvic M., Ysabel C. Casart, and María I. Camejo. "Nitric oxide and oxidative stress in placental explant cultures." Systems Biology in Reproductive Medicine 62, no. 1 (September 14, 2015): 11–16. http://dx.doi.org/10.3109/19396368.2015.1075163.

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47

Donovan, Stacy L., and Michael A. Dyer. "Preparation and square wave electroporation of retinal explant cultures." Nature Protocols 1, no. 6 (December 2006): 2710–18. http://dx.doi.org/10.1038/nprot.2006.454.

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48

Do, Vi Nguyen Tuong, Shan-Te Hsu, and Yung-I. Lee. "Clonal Propagation In Vitro of Paphiopedilum Hybrids from Adult Plants." HortScience 54, no. 9 (September 2019): 1565–69. http://dx.doi.org/10.21273/hortsci13791-18.

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The aim of this study was to develop an efficient protocol for shoot tip culture from adult plants of Paphiopedilum Pfitzer. A considerable seasonal effect on explant collection was observed in the aseptic cultures established from adult plants, including the survival and microbial contamination of explants. The shoot tip explants excised from adult plants in February and May showed higher survival and had less contamination than those explants excised in August and November. Moreover, the season of explant collection also affected the subsequent shoot forming capacity and multiplication of axillary buds. In Paphiopedilum ‘In-Charm Silver Bell’, higher shoot forming capacity was observed in February and May, whereas higher shoot multiplication was observed only in February. In Paphiopedilum ‘Hsinying Maudiae Leopard’, both February and May were optimal timing for shoot forming capacity and multiplication. We also demonstrated the effectiveness of transcinnamic acid (tCA), an antiauxin chemical in diminishing the apical dominance of shoot tip explant and thus improving the axillary bud outgrowth. In P. ‘In-Charm Silver Bell’, the addition of 100 μM tCA plus 13.3 μM 6-benzylaminopurine (BA) for 1 month promoted axillary shoot bud formation from shoot tip explants as compared with the control.
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49

Emoghene, B. O., M. Idu, C. R. Eke, and O. Asemota. "Effects Of Different Sterilization Regimes & Growth Regulators On Micropropagation Of Female Date Palm (Phoenix dactylifera L.)." Nigerian Journal of Biotechnology 37, no. 1 (August 28, 2020): 159–68. http://dx.doi.org/10.4314/njb.v37i1.17.

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The success of in vitro culture techniques is always hampered by microbial contamination. The present study was carried out to develop an efficient protocol for date palm explants sterilization for successful somatic embryos induction and plantlets formation of some date palm varieties. The shoot tips were treated with different sterilizing agents at different concentrations and durations of exposure. The use of ethanol (70%), sodium hypochlorite (3.5% & 70%) and mercuric chloride (0.2%) with or without addition of Tween-20 had different effects on decontamination of the date palm explants. The percentage of explants contaminated with bacteria for sterilants 1, 2 and 4 was 18.8%, 6.3% and 6.3% respectively while 25%, 37.5%, 31.25% and 6.25% were contaminated with fungi for sterilants 1, 2, 3 and 4 respectively. Under the conditions used, a combination of antioxidants (Citric and Ascorbic acids at 100mg/l), 0.2% mercuric chloride and 3.5% sodium hypochlorite solution with 3 drops/100ml of Tween-20 helped in the reduction of chlorosis, contamination and die-back in the shoot tip explants. The explants were further cultured in appropriate media for callus initiation and subsequent somatic embryo induction. Optimal embryogenic callus was obtained from the shoot explant of sterilant number 4 which had the minimal contamination and die-back of all the cultures. After 3 subcultures, the somatic embryos formed were multiplied for shoot development. From this study, we established that the use of appropriate surface sterilant at suitable concentration and duration of exposure of date palm explant to it is indispensable for maximum responses of in vitro cultures. Keywords: Date palm, Microbial contamination, Sterilizing agents, in vitro, Somatic embryos
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50

Khan, Sagib B., John E. Preece, Bradley H. Taylor, and J. W. Van Sambeek. "Response of Adult Black Walnut (Juglans nigra L.) Nodal Explants on DKW and LP Media." HortScience 30, no. 4 (July 1995): 874B—874. http://dx.doi.org/10.21273/hortsci.30.4.874b.

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Abstract:
Branch tips (30 to 40 cm long) of adult black walnut were forced in a half-strength solution of Long and Preece medium (LP) salts (minus iron) plus 1 mM 8-hydroxyquinoline citrate (8-HQC). The resulting softwood shoots were surface-disinfested and cut into 1.5-cm-long nodal segments. Explants were placed on two media: Driver and Kuniyuki Walnut medium (DKW) or LP with four plant growth regulator combinations: 5 μM BA with 0.05 μM IBA, 10 μM BA, 1 nM TDZ, or 10 nM TDZ in a factorial arrangement. Gelrite was used as the gelling agent. Explants were transferred to fresh medium on days 1, 3, 5, and 7 after initiation, then weekly. Data recorded 60 days after culture initiation showed more and longer shoots and leaves, greater explant diameter, more green (living) tissue, and less exudation per explant on LP than on DKW. Greatest explant and shoot length were observed when the medium contained 10 nM TDZ. BA (10 μM) and LP were best for long-term maintenance of cultures
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