正常小鼠阴道上皮细胞体外原代培养探索

Primary culture of normal mouse vaginal epithelial cells in vitro

探索正常小鼠阴道上皮细胞体外原代培养技术,在短时间内获取纯度高、活性好的阴道上皮细胞,为后续更好地进行女性生殖道微生态失常引起的相关疾病的研究奠定基础。利用中性蛋白酶和胰酶分离正常小鼠阴道上皮细胞,在Eplife培养基的基础上添加了氯化钙来培养,用光镜、电镜来观察细胞生长情况,免疫组化技术鉴定细胞来源。光镜、电镜下可见体外阴道上皮细胞呈铺路石状分布,纯度高,可传3~4代,P-CK、CK5/6角蛋白单克隆抗体免疫组化染色阳性。正常小鼠阴道上皮细胞体外原代培养、传代成功,该方法行之有效。

To explore the technique of primary culture of normal mouse vaginal epithelial cells in vitro,obtain high pureness and activity vaginal epithelial cells within a short time,and make foundation to doing further research in the reproductive tract diseases caused by the unbalanced of the virginal micro-ecoenvironment.Mouse vaginal epithelial cells were cultured in vitro with Epilife,after being isolated by DispaseⅡand Trypsin.On the basis of Eplife medium,calcium chloride was added to culture medium,cells growth were observed by inverted microscope and confocal microscope,and the origin of cells was identified by immuno-histochemical technique.The purity is high and can be passaged 3~4 times.Cells looked like a typical cobblestone under inverted microscope and confocal microscope.While P-CK,CK5/6 immuno-histochemical staining were positive.The primary culture of normal mouse vaginal epithelial cells in vitro issuccessful and the method is effective.