中文摘要：

目的：探讨采用无血清培养基（Ker—SFM）原代培养成人宫颈上皮细胞的效果和影响因素。方法：取65例因子宫良性病变切除子宫患者的宫颈组织，采用diapaselI和胰蛋白酶一EDTA消化法获得宫颈上皮细胞悬液，于上皮细胞专用Ker—SFM中培养，采用免疫细胞化学法测定细胞中E一钙黏蛋白（E—cadherin）及波形蛋白（vimentin）的表达，对细胞纯度进行鉴定，并用透射电镜及扫描电镜观察细胞形态。结果：培养成功率69．23％（45／65），原代培养12～15天细胞生长达80％以上，早期细胞生长状态良好，传代至3代，细胞出现生长衰退。细胞鉴定E—cadherin阳性，vimentin染色阴性，电镜观察细胞符合上皮细胞特点。影响培养成功率的因素主要包括患者年龄大小、取材处组织有无损伤、子宫离体时间、术前是否有阴道或宫颈炎症以及培养基的选择。结论：采用酶消化法及Ker—SFM培养可获得纯度较高的成人宫颈上皮细胞，可作为宫颈癌基础研究的实验模型。

英文摘要：

Objective： To explore the effect of primary culture of human cervical epithelial cells with serum - free medium （ Ker, SFM） and influencing factors. Methods： The cervical specimens from 65 patients after excision of uterine benign lesions were selected, suspension of cervical epithelial cells were obtained by diapase II and trypsin - EDTA digestion method, and then they were cultured in Ker - SFM special for epithelial cells ; immunocytochemical method was used to detect the expressions of E - cadherin and vimentin; the purity was determined, transmission electron microscopy and scanning electron microscopy were used to observe cell morphology. Results： The suc- cessful rate of primary culture was 69. 23% （45/65）, the growth rate was more than 80% after 12 - 15 days, the growth of cells during early phase was good, after passaging to the third generation, grown recession was found; E - cadherin positive staining and vimentin negative stai- ning were determined; the cells under electron microscope accorded with the characteristics of epithelial cells ; the influencing factors of suc- cessful rate of primary culture mainly included age, cervical injury or not where drawing tissues, excision of uterine, preoperative vaginitis or cervicitis, and choice of culture medium. Conclusion： Human cervical epithelial cells with high purify can be obtained by enzyme digestion method and Ker - SFM culture, which can be used as an experimental model for basic studies of cervical cancer.