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Postmortem recovery of proliferative fibroblasts up to three days in livestock ear skin stored at 35°C

机译:储存在35°C的牲畜耳朵皮肤中的增殖性成纤维细胞的死后恢复长达三天

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Effect of elevated temperature (35°C) on postmortem survival of cells in goat, sheep and bovine tissues was studied. Skin explants (n = 30; 2-3 mm2) were cultured in petri dishes after 2, 24, 48, 72, 96 and 120 hours of postmortem interval in DMEM media supplemented with 10% FBS, 50 units/mL of penicillin, 50 μg/mL of streptomycin and 2.5 μg/mL of fungizone. Outgrowth of cells around the explants was observed up to 72 h in sheep, 48 h in bovine and 24 h in goats. In general, the number of explants exhibiting outgrowth as well as the level of confluence decreased with increasing postmortem time interval. Secondary cultures established from primary cells for 72 h postmortem interval show cytogenetically stable chromosomes with 54XX[17] normal female sheep karyotype, comparable cell morphology, and growth curve to that of fresh tissue derived cells. Till date these cells have been passaged 21 times and show normal growth. These results suggest that normal, proliferative cells can be recovered from skin tissues stored at an elevated temperature of 35°C in livestock from 24-72 h of postmortem interval. Reprogramming of these cells to clone the dead animals or their use for cell therapies remain to be seen in future.
机译:研究了高温(35°C)对山羊,绵羊和牛组织中细胞的死后存活的影响。死后间隔2、24、48、72、96和120小时后,在补充有10%FBS,50单位/ mL青霉素,50的DMEM培养基中,在皮氏培养皿中培养皮肤外植体(n = 30; 2-3 mm2) μg/ mL的链霉素和2.5μg/ mL的真菌区。在绵羊中长达72小时,在牛中为48小时,在山羊中为24小时,观察到外植体周围细胞的生长。通常,随着死后时间间隔的增加,表现出外植体的外植体数量以及融合水平降低。在死后72小时内从原代细胞建立的次要培养物显示出具有54XX [17]正常雌性羊核型的细胞遗传学稳定染色体,具有与新鲜组织衍生细胞相当的细胞形态和生长曲线。到目前为止,这些细胞已经传代了21次并显示出正常的生长。这些结果表明,从死后间隔的24-72小时,可以从储存在牲畜中35℃的高温下的皮肤组织中恢复正常的增殖细胞。这些细胞的重编程以克隆死亡的动物或将其用于细胞治疗仍有待观察。

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