改进的酶消化法培养新生大鼠Müller细胞

Modified enzyme-digesting method for culturing Muller cells of newborn-rat

目的通过改进的酶消化法,简化Muller细胞的培养和纯化过程。方法采用将眼球培养前处理和机械吹打、贴壁后反复胰蛋白酶消化的方法培养新生大鼠Muller细胞,进行免疫组织化学染色和光电镜观察。结果培养的Muller细胞光镜下胞体较大,胞浆丰富,电镜下细胞器丰富,可见糖原和直径为10nm的微丝,免疫组织化学示95%以上细胞谷氨酰胺合成酶阳性,神经胶质纤维酸性蛋白阴性。结论改进的酶消化法是一种简单可行的Muller细胞培养方法。

Objective To simplify the procedure of culture and purification of Muller cells by an modified enzyme-digesting method. Methods Eyeball of newborn rat was pretreated and mechanically dissociated and repeated enzyme-digesting after cells anchor for the culture of Miiller cell. Cultured-Muller cell was identified by immunocytochemistry, and the morphology of cells was observed by inverse phase microscope and electronic microscope. Results Cultured-Muller cells had large cell body and relatively wide cytoplasmic expansion, and glycogen and intermediate filaments could be observed in cytoplasm. More than 95% cells were positive staining for glutamine synthetase and negative for glial fibrillary acidic protein. Conclusion Modified enzyme-digesting method is a simple and practical way for culturing Miiller cells.