大鼠尾芽胚视杯干细胞的分布与特征

Distribution and features of the optic cup stem cells in embryonic rat at tailbud stage

目的 探索视杯干细胞在大鼠尾芽胚(第12 5天胚龄)的分布与特征。 方法 采用免疫组织化学技术,检测视杯干细胞在大鼠尾芽胚(第12 .5天胚龄)视杯组织中的分布;分离视杯细胞,体外无血清培养,应用免疫细胞化学技术分析其增生能力以及血清诱导分化前后CHX10和多种成熟视网膜细胞特异性标记蛋白的表达,以了解这一发育时期视杯组织的分化特点。 结果 大鼠尾芽胚(第12 .5天胚龄)的视杯干细胞主要分布在视杯的内外层和边缘层,不表达成熟视网膜细胞特异性标记蛋白。从尾芽胚视杯中分离出的细胞具有单细胞克隆能力,CHX10表达阳性,血清诱导后表达多种成熟视网膜细胞特异性标记蛋白:Thy1.1、神经胶质纤维酸性蛋白(GFAP)、蛋白激酶C(PKC)α和rhodopsin。 结论 大鼠尾芽胚(第12 .5天胚龄)视杯主要由未分化的细胞组成,视杯干细胞的分布集中在视杯内层和边缘层。体外培养的视杯干细胞增生能力强,经诱导分化后表达多种成熟视网膜细胞特异性标记蛋白。

Objective To explore the distribution and features of the optic cup stem cells in embryonic rat at tailbud stage. Methods The distribution of optic cup stem cells in optic cup tissue in 12.5-embryonic-day-old rats was observed by immunohistochemistry. The separated cells from optic cup were cultured with serum-free media, and immunofluorescence technique was used to detect the ability of hyperplasia of stem cells and expression of CHX10 antigen and specific antigens of mature retinal cells before and after differentiation. Results The optic cup stem cells in embryonic rat at tailbud stage were mainly located at inner, outer, and marginal layer of optic cup. No expression of specifically marked protein of mature retinal cells was detected. The cells separated from optic cup had the ability of single-cell clone, positive expression of CHX10 and expression of several specific antigens of mature retinal cells after the inducement, including Thy1.1, glial fibrillary acidic protein (GFAP), protein kinase C (PKC) α, and rhodopsin. Conclusion Optic cup of 12.5-embryonic-day-old rats composes of undifferentiated cells, and the stem cells are mainly located in optic cup inner and marginal. High ability of hyperplasia of the optic cup stem cells cultured in vitro is found. The cells, which are retinal stem cells, can express several specifically marked proteins of mature retinal cells after inducement and differentiation.