膀胱上皮细胞的体外原代培养方法

背景:国内外获得原代人正常膀胱上皮细胞的方法包括酶消化培养法、刮削培养法、组织块培养法,这些方法各有优缺点,培养基中成分多不易把握,效率不高。目的:分析人膀胱上皮细胞原代培养的最佳方法。方法:在同一实验条件下,采用酶消化培养法、刮削培养法、组织块培养法3种不同的原代细胞培养方法培养人正常膀胱黏膜膀胱上皮细胞。结果与结论:酶消化培养法、刮削培养法、组织块培养法各组膀胱上皮细胞培养成功率分别为13.3%,26.7%,86.7%,3组间比较差异有非常显著性意义(P<0.001)。3组培养的人膀胱上皮细胞角蛋白AE1/AE3荧光染色均呈阳性。说明组织块培养法是一种简单,短期内可扩增出较多纯净的膀胱上皮细胞的原代培养方法。

Primary culture of bladder epithelial cells in vitro

BACKGROUND:There are three methods to obtain human bladder epithelial cells:enzymatic digestion,scraping and tissue plant.Each of the three methods has its advantage and disadvantage.There are too many ingredients in the culture medium to deal with,besides,the efficiency is low.OBJECTIVE:To approach the best method for primary culture of human bladder epithelial cells.METHODS:Human bladder epithelial cells from normal bladder mucosa were cultured with three different primary cell culture methods:enzymatic digestion,scraping and tissue plant.RESULTS AND CONCLUSION:The successful rates of bladder epithelial cells cultured with enzymatic digestion,scraping and tissue plant were 13.3%,26.7% and 86.7% respectively,and the differences in successful rates were significant(P < 0.001).Fluorescent staining with AE1/AE3 antibody confirmed that the cultured bladder epithelial cells were positive for AE1/AE3 epithelial cytokeratins.Tissue plant is a simple and feasible primary culture method for bladder epithelial cells culture,and a great amount of pure bladder epithelial cells can be obtained in a short time using this method.