人角膜缘干细胞体外培养后生物学特性的变化

目的 探讨人角膜缘干细胞体外培养后细胞形态学，抗原性和增殖能力的变化。方法 消化培养法体外培养人角膜缘干细胞。获得细胞单层并观察细胞形态学变化。利用间接免疫荧光组化检测人角膜缘组织HLA-DR抗原在培养前，后的变化，检测培养后细胞增殖核抗原和角膜上皮64KD角蛋白的表达。结果 原代细胞在培养1周形成单层细胞形态以圆柱状细胞为主，培养前角膜缘上皮下少星HLA-DR抗原分布，培养后单层细胞DR抗原表达阴性；单层细胞中多量，散在分布的增殖细胞核抗原阳性细胞，并且部分细胞表达64KD角蛋白，结论 角膜缘干细胞体外培养后分化为角膜上皮，不表达HLA-DR抗原，介仍保持较强的分裂增殖能力，可能是临床移植治疗干细胞缺乏眼表疾病患者的一种有效手段。

Biological characteristics changes of human limbal stem cells after primary culture in vitro

Objective To observe the changes of limbal stem cell morphology?antigen and proliferative ability after primary culture in vitro.Methods Human limbus was cultured by enzymatic digestion culture method and the morphology of the monolayer cultured cell was observed. The expression of HLA-DR antigen?proliferative cell nuclear antigen(PCNA) and 64KD keratin at limbus and cultured cells were detected by indirect immunochemical staining. Results Eighty percent confluence was reached at about 1 week, mainly consisting of basal columned cells. HLA-DR antigen was positive subepithelium at limbus, but negative in cultured cells; The staining of PCNA and 64KD keratin were positive in cultured cells. Conclusion The cultured epithelium cells which are taked from limbus tissue can be differentiated into corneal epithelium, without HLA-DR expression but with high proliferative potency. These cultured epithelium may be useful for transplantation to cure ocular surface diseases.