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目的探讨人正常腹膜间皮细胞原代培养方法。方法以胰酶和乙二胺四乙酸磁性搅拌分离人正常腹膜间皮细胞,倒置相差显微镜观察细胞形态结构和大体生长过程,Calretinin免疫荧光染色鉴定间皮细胞,初步观察传代细胞的贴壁生长情况。结果培养第4天细胞贴壁良好,第15天呈铺路石状、生长良好,1个月时细胞密集铺满皿壁;腹膜间皮细胞Calretinin表达都呈阳性;传代腹膜间皮细胞数量显著减少。结论磁性搅拌酶分离培养可得到数量多、纯度高的原代人正常腹膜间皮细胞,能够满足一般实验要求。
Objective To investigate the method of primary culture of normal human peritoneal mesothelial cells. Methods Human normal peritoneal mesothelial cells were isolated by magnetic stirring with trypsin and ethylenediaminetetraacetic acid. Morphology and growth were observed by inverted phase contrast microscope. The mesothelial cells were identified by Calpactin immunofluorescence staining. The adherent growth of subcultured cells was observed . Results On day 4, the cells adhered well, and became paved stone on the 15th day. The cells grew well at 1 month. The cells were densely covered with the wall of the dish. The expression of Calretinin in the peritoneal mesothelial cells was positive. The number of passage peritoneal mesothelial cells was significantly decreased . Conclusion Isolation and culture of magnetic agitation enzyme can obtain a large number of primary human normal peritoneal mesothelial cells with high purity, which can meet the general experimental requirements.