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目的 比较表皮干细胞(KSC)在两种体外培养体系中的生长增殖状况。方法 用差速粘附法分离出的KSC,分别置于无饲养层、无血清培养基以及有饲养层、有血清培养基的培养条件下培养。比较KSC在两种培养体系中的生长增殖、克隆形成率、生长曲线和KSC相关标记物的表达等指标。结果 在第二种培养条件下,KSC在生长过程中不存在明显的接触抑制,在较长的体外培养时间内始终保持较高的增殖潜能。两种培养体系间KSC的克隆形成率有显著差异(P<0.01)。流式细胞仪检测两种培养体系间整合素β1阳性率无显著差异,而强阳性率则有显著性差异(P<0.05)。结论 体外培养和扩增KSC时,使用人成纤维制备的饲养层和含胎牛血清的培养基,较无饲养层、无血清的培养基,更有利于KSC的扩增及其表型的维持。
Objective To compare the growth and proliferation of epidermal stem cells (KSC) in two in vitro culture systems. Methods KSCs isolated by differential adhesion method were cultured in feeder-free, serum-free medium and feeder layer with serum-containing culture medium. The growth and proliferation, clone formation rate, growth curve and expression of KSC related markers of KSC in two culture systems were compared. Results Under the second culture condition, KSC did not show obvious contact inhibition during growth, and maintained a high proliferative potential over a longer period of in vitro culture. The clonogenic rates of KSC between the two culture systems were significantly different (P <0.01). Flow cytometry showed no significant difference in the positive rate of integrin β1 between the two culture systems, but the strong positive rate was significantly different (P <0.05). Conclusion In vitro culture and expansion of KSC, the culture medium containing human Fibroblasts and medium containing fetal bovine serum is more conducive to the expansion of KSC and the maintenance of its phenotype compared with the culture medium without serum or serum. .