改良筛网法分离培养人子宫内膜细胞  

A modified filter method to isolate and culture human endometrial cell

作　　者：陈淑琴[1] 金文艳[1] 黄佳明[1] 姜红叶[1] 姚书忠[1] 

机构地区：[1]中山大学附属第一医院妇产科,广州510080

出　　处：《现代妇产科进展》2012年第4期280-285,共6页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金面上项目(No:81070472)

摘　　要：目的:探索一种能获得较高纯度和较多数量子宫内膜细胞的分离培养方法。方法:取12例患者各1mg新鲜子宫内膜组织,经0.25%Ⅰ型胶原酶(含有0.005%DNA酶)消化后,将所得细胞悬液一分为二,分别采用传统筛网法和改良筛网法(二次筛网过滤和低速离心相结合)分离并记录所得细胞数;体外培养后采用光学显微镜观察,用免疫细胞化学和免疫荧光化学方法鉴定纯度,比较两种方法分离子宫内膜细胞的纯度和数量。结果:两种方法均有11例组织分离培养成功。传统筛网法分离的子宫内膜腺上皮细胞和间质细胞的纯度分别为(89.54±3.75)%和(92.82±1.60)%,细胞数分别为(0.47±0.09)×105个和(23.73±5.37)×105个。改良筛网法分离的腺上皮细胞和间质细胞纯度分别为(91.95±0.47)%和(99.28±0.21)%,细胞数分别为(8.09±1.24)×105个和(27.23±0.50)×105个。改良筛网法分离的腺上皮细胞数量和间质细胞纯度均明显高于传统筛网法,有统计学差异(P均=0.000)。结论:改良筛网法操作简单,能获得较高纯度的子宫内膜间质细胞及较多的子宫内膜腺上皮细胞。Objective：To establish a modified method to isolate endometrial glandular epithelial cells and stromal cells with higher purity and greater number in vitro.Methods：The fresh endometrium of 1 mg from 12 endometrial specimens were obtained,digested with 0.25% collagenaseⅠ containing 0.005% DNA enzyme and then seperated by filter method and modified method which refers to combination serial filtration with gradient centrifugation,respectively.The cell number was record after separation by cell count.The purity of the isolated endometrial glandular epithelial cells and stromal cells were identified morphologically by light microscopy,and stained for vimentin and cytokeratins by immunohistochemistry and immumofluorescent assay.Results：The cells were isolated and cultured from 11 endometrial specimens successfully with both methods.（8.09 ± 1.24）× 105 epithelial cells and（27.23 ± 0.50）× 105 stromal cells with the purities of（91.95 ± 0.47）% and（99.28 ± 0.21）% respectively were isolated from endometrial tissues using the modified method,whereas（0.47 ± 0.09）× 105 epithelial cells and（23.73 ± 5.37）× 105 stromal cells with the purities of（89.54 ± 3.75）% and（92.82 ± 1.60）% respectively were obtained by filter method.Compared to previous filter methods,the purity of glandular epithelial cells and cell count of stromal cells by modified method were significantly higher than those by filter method,respectively（P= 0.000）.Conclusion：We have developed a modified method to isolate endometrial glandular epithelial and stromal cell populations with a higher purity and greater number in vitro.

关 键 词：子宫内膜 腺上皮细胞 间质细胞 细胞培养 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]