组织块法和酶消化法体外培养原代鼠阴道上皮细胞的研究

Compariation of Tissue Explant and Enzyme Digestion Method for Rat Vaginal Epithelial Cells in vitro

为探讨更佳的阴道上皮细胞体外培养技术,为组织工程化阴道动物模型提供种子细胞,分别应用组织块法和酶消化法原代培养大鼠阴道上皮细胞,观察两种方法细胞生长所需时间、细胞形态和生长特性,免疫组化进行鉴定。结果表明,两种细胞培养方法均能获得不规则圆形或多边形的阴道上皮细胞,其传代后增殖特性和生长曲线基本一致,角蛋白染色阳性,但酶消化法较组织块法细胞贴壁快、生长时间短、产量大、细胞纯度高、能较迅速获得较多细胞用于组织工程阴道的构建。

Vaginal epithelial cells from Sprague-Dawley rats were primary cultured by tissue explant and enzyme digestion method respectively to explore the better culture technique in vitro and provide seed cells for engineering of vaginal tissue.Time of primary culture,the cell morphology and proliferation characteristics were oberserved under light microscopic.The cells were identified with P-CK anti by immunohistochemistry staining. The results indicated that the vaginal epithelial cells which were obtained by the two methods were polygon-like or irregular globular,and their morphology and growth characteristics were essentially consistent.The expression of pan-cytokeratin staining of the cultured cells were positive.However,compared with explant method,the culture cells using enzyme digestion method displayed more faster adhesion,shorter growth time,and higher cells productivity and purity.The larger number of epithelial cells were rapidly obtained to construct engineering of vaginal tissue by enzyme digestion method.