中性蛋白酶和胰酶分离阴道黏膜上皮细胞的对比研究

Isolation of Vaginal Mucosal Keratinocytes:a Comparative Study of Dispase and Trypsin

目的:建立新的阴道黏膜上皮细胞分离方法,体外培养阴道黏膜细胞供进一步研究。方法:利用中性蛋白酶(dispase)和胰酶分离人阴道黏膜上皮细胞,并通过形态学、角蛋白13(CK13)染色、细胞产量和克隆形成率等比较分离效果。结果:dispase组的细胞产量及克隆形成率更高。形态学和CK13染色均可见胰酶的分离作用并不完整,细胞大部分仍附于固有层,而dispase组基底细胞则完整地分离开。结论:酶消化法能成功分离阴道黏膜细胞,在体外生长良好,而dispase分离效果更好。

Objective： To present a new method for isolating vaginal mucosal keratinocytes, and culturing them in vitro for further study. Methods： Keratinocytes from human vaginal mucosa were isolated using either dispase or trypsin, and their purity and viability were assessed by histology, cytokeratin 13 （CK13） staining, cell yield and colony-forming efficiency. Results： The cell yield and colony-forming efficiency were higher in the cells isolated using dispase. Histological examination and CK13 staining showed that treament with trypsin resulted in incomplete separation of the cells, with large numbers of cells remaining attached to the lamina propria. In contrast, basal cells were separated completely when dispase was used for isolation. Conclusion： Vaginal mucosal keratinocytes can be isolated using proteases, and can be successfully grown in vitro. Although tyrpsin can be used to isolate the cells, the use of dispase results in better isolation and separation from the lamina propria, and better cell viability during culture.