优化胰蛋白酶消化强度进行人子宫内膜上皮细胞培养技术探讨  

Study on culture technology of human endometrial epithelial cells by optimizing trypsin digestion intensity

作　　者：谢伟[1] 秦雯[2] 蒋凌云[1] 莫馥华 朱时敏[3] 顾克英 罗远富 覃爱平[1] 

机构地区：[1]广西医科大学第一附属医院医学生殖中心,广西南宁530021 [2]广西医科大学第一附属医院病理科,广西南宁530021 [3]广西医科大学,广西南宁530021

出　　处：《现代医药卫生》2016年第11期1606-1608,1611,共4页Journal of Modern Medicine & Health

基　　金：广西优秀博士论文培养计划资助项目(YCBZ2014033)

摘　　要：目的探讨优化改良的人子宫内膜原代上皮细胞培养技术。方法将2014年7月1日至2014年9月30日需行诊断性清宫术的患者21例分为0.125%胰蛋白酶组（A组）、0.250%胰蛋白酶组（B组）和1 mg/mL胶原酶组（C组）。同时结合使用100目滤网（150μm）及400目（38μm）滤网进行子宫内膜上皮及间质细胞分离,比较各组间子宫内膜上皮细胞纯度和生长状态。结果子宫内膜细胞培养成功19例,A组子宫内膜组织污染1例,C组因细胞量过少接种失败1例。细胞获得后培养进行上皮细胞团百分比计算,A组为（85.71±1.38）%、B组为（75.28±1.50）%和C组为（84.86±2.34）%,其中A组和C组的上皮细胞百分比高于B组,差异均有统计学意义（P〈0.05）。细胞培养3~5 d后待细胞基本铺满培养孔,免疫组化辣根过氧化物酶底物显色（ABC法）进行纯度检测,A组为（90.28±2.13）%、B组为（84.57±2.76）%和C组为（86.14±4.06）%,A组细胞纯度均大于B、C组,差异均有统计学意义（P〈0.05）。A组子宫内膜上皮细胞获得数量和纯度均高于其他两组,差异均有统计学意义（P〈0.05）。结论 0.125%胰蛋白酶消化子宫内膜组织结合100目滤网（150μm）及400目（38μm）滤网过滤,可以稳定地获得质量良好的子宫内膜上皮原代细胞。Objective To investigate an optimized and improved culture technology of primary human endometrial ep-ithelial cells. Methods Twenty-one patients undergoing diagnostic uterine curettage from July 1 to September 30, 2014 were divided into the 0.125% trypsin group（A）,0.250% trypsin group（B）and 1mg/m L collagenase group（C）. Meanwhile 100-mesh strainer（150 μm） and 400-mesh strainer（38 μm） were used to separate the endometrial epithelial cells and interstitial cells. Then the cells purity and growth state were compared among various groups. Results Nineteen cases succeeded in the endometrial epithelial cell culture. One case in the group A was contaminated by endometrial tissues,1 case in the group C failed in the inoculation due to too little cell amount. In conducting the epithelial cell cluster percentage calculation after obtaining cells and inoculat-ing,the group A was（85.71±1.38）%,group B was（75.28±1.50）% and group C was（84.86±2.34）%,in which the percentage of epithelial cells in the group A and C was higher than that in the group B,and the difference was statistically significant（P 〈0.05）. After the cells basically overspread the culture holes by 3-5 d of cell culture,the purity was detected by using the immunohistochemistry ABC method,the group A was（90.28±2.13）%,group B was（84.57±2.76）% and group C was（86.14±4.06）%. The purity of the group A was higher than that of the group B and C,and the difference had statistic significance（P 〈0.05）. The acquired endometrial epithelial cells amount and purity in the group A were higher than those in the other two groups,the difference was statistically significant（P 〈0.05）. Conclusion 0.125% trypsin digesting endometrial tissues and combining with the filtration by100-mesh strainer（150 μm） and 400-mesh strainer（38 μm）can stably acquire good-quality primary endometrial epithelial cells.

关 键 词：子宫内膜 上皮细胞 细胞培养技术 胰蛋白酶 

分 类 号：R339.22[医药卫生—人体生理学]