组织块联合胰酶消化法培养兔角膜缘干细胞的初步探索

Preliminary exploration of rabbit corneal limbus stem cells cultured with tissue block and enzyme digestion method

目的:建立一种简便、高效的兔角膜缘干细胞原代培养方法。方法:获取兔角膜缘组织,采用组织块联合胰蛋白酶消化法进行兔角膜缘干细胞体外培养,倒置显微镜下观察细胞生长情况,HE染色观察细胞形态和结构特点,并运用免疫组织化学技术进行细胞鉴定。结果:采用组织块联合胰蛋白酶消化法可以简便、快速地培养出兔角膜缘干细胞,显微镜下动态观察细胞生长良好,有较高的增殖能力;HE染色证实细胞形态和结构正常;AE5及P63免疫组织化学鉴定呈阳性。结论:采用组织块联合胰蛋白酶消化共同培养的方法,建立了一种简便、高效的兔角膜缘干细胞原代培养模式。

AIM:To establish a simple and efficient method for the primary culture of rabbit corneal limbus stem cells.METHODS:Obtained the limbal tissues from rabbits,used tissue block and enzyme digestion method to culture the corneal limbus stem cells in vitro.The growth characteristics of the cultured cells in vitro were observed under inverted microscope.By means of HE,the morphology and construction features of cells were observed.And immunohistochemical method was used to identify the cultured cells.RESULTS:Rabbit corneal limbus stem cells could be fast and simply cultured by using tissue block and enzyme digestion method.The dynamic observation under microscope showed that rabbit corneal limbus stem cells grew well with a higher proliferative capacity.In HE staining,the morphology and structure of cells were normal.AE5 and P63 cellular immune identification were positive.CONCLUSION:Tissue block and enzyme digestion method could be a simple and efficient mode for the primary culture of rabbit corneal limbus stem cells.