摘要
目的了解角膜缘干细胞体外培养的增殖分化规律。方法组织块法培养细胞,测定细胞克隆形成率(CFE),免疫荧光染色检测干细胞表达角质蛋白K3的状况。结果原代培养21天左右细胞生长达到饱和,传第1代7~10天形成单层,CFE为9.52%±4.97%;传第2代7天,CFE为4.25%±2.10%(P<0.01)。正常角膜缘基底细胞不表达角质蛋白K3;原代培养的干细胞亦不表达K3,传第1代细胞有部分表达。结论人角膜干细胞位于角膜缘基底部,培养的角膜缘干细胞早期具有较高的增殖力并保持干细胞的分化特性。
ObjectiveTo examine the proliferation and differentiation of human limbal stem cells in vitro.MethodsLimbal stem cells were cultured by tissue patch and examined with colony forming efficiency(CFE)and expression of keratin 3 in normal corneal tissue and cultured corneal epithelial cells on coverslip by immunofluorescent staining.ResultsThe limbal stem cells reached confluence at 21d after seeded in primary culture and 7~10d in first passage and 7d in second passage.The means CFE of stem cells (SCs)were 9.52%±4.97% in first passage and 4.25%±2.10% in second passage ( P <0.01).Human limbal epithelial cells did not expressed K3 except superficial cells.And human limbal stem cells also did not expressed K3 in primary cultured but partly expressed in first passaged cells.ConclusionHuman corneal SCs are part of limbal basal epithelial cells.The SCs in vitro in primary and first passsage have higher proliferating capacity and maintain characteristics of SCs.
出处
《眼科研究》
CSCD
1999年第2期81-83,共3页
Chinese Ophthalmic Research
关键词
角膜缘干细胞
细胞增殖
细胞分化
体外培养
uman limbal stem cells proliferation and differentiation in vitro