鲍曼囊壁层上皮细胞培养与鉴定

Chinese Journal of Nephrology(2022)

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Abstract
目的:分离和培养小鼠鲍曼囊壁层上皮细胞,为进一步深入研究其生理作用提供基础。方法:通过细胞筛联合磁性分离的方法分离小鼠肾小体,原代培养后获取壁层上皮细胞,诱导壁层上皮细胞分化为足细胞。使用免疫荧光染色、实时定量PCR和Western印迹法检测壁层上皮细胞和足细胞特异性标志物。结果:原代培养的壁层上皮细胞呈现铺路石样,表达壁层上皮细胞特异性标志物Claudin-1和干细胞标志物CD133和CD24,不表达肾小管上皮细胞蛋白、系膜细胞和足细胞标志蛋白。体外培养传代至6代的壁层上皮细胞仍表达Claudin-1、CD133和CD24。经过诱导分化后,壁层上皮细胞可表达足细胞特异蛋白WT-1和Synaptopodin。结论:细胞筛联合磁性分离法提取肾小体,体外培养获得的小鼠壁层上皮细胞可以在体外诱导表达足细胞标志蛋白。
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Key words
Nephrons,Epithelial cells,Cell separation,Cells, cultured,Identification
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