中文摘要：

目的：研究大鼠阴道黏膜上皮细胞（RVECs）体外培养的影响因素,探索建立RVECs稳定的体外分离培养方法。方法：比较大鼠动情周期、DispaseⅡ配制方法、Dis-paseⅡ浓度、胰酶消化法及培养基对RVECs体外培养的影响。结果：动情前期的大鼠阴道上皮层更厚,消化后可获得更多RVECs;上皮层用D-KSFM配制的DispaseⅡ消化后能很好的贴壁,用PBS配制的DispaseⅡ消化后细胞贴壁欠佳;1.2U/ml DispaseⅡ的分离效果优于0.6U/ml DispaseⅡ;三步胰酶消化法比一步胰酶消化法可获得更多的贴壁细胞;RVECs在含血清培养基中生长快,但易受成纤维细胞污染。结论：动情前期的大鼠阴道组织依次经D-KSFM配制的1.2U/ml DispaseⅡ及三步胰酶消化后,在不含血清的D-KS-FM中培养,可获得更多的RVECs,细胞显示上皮细胞特征。

英文摘要：

Objective：To explore factors impacting on primary culture of rat vaginal epithelial cells（RVECs） in vitro,and establish a reproducible protocol for RVECs separation and culture that could be applied in tissue engineering.Method：Comparing influence of rats in different estrus,Dispase Ⅱ preparation,Dispase Ⅱ concentration,trypsine digestion and culture medium on RVECs culture.Result：Epithelial layer of rat in preoestrus was thicker and more abundant than other phases.More RVECs could attach after digestion with Dispase Ⅱ solving in D-KSFM than in PBS.Separation of epithelium from underlying connective tissues with 1.2U/ml Dispase was more complete than that of 0.6U/ml Dispase.More RVECs could attach after digestion with trypsine by 3 steps than that by one step.RVECs grew fast in medium containing fetal bovine serum,but was vulnerable to fibroblast cells contamination.Conclusion：The vaginal tissue of rat in preoestrus digest with 1.2 U/ml Dispase Ⅱsolving in D-KSFM and with trypsine by 3 steps in succession,then culture in the D-KSFM without FBS,more RVECs demonstrating epithelial characterization can be obtained.