子宫内膜异位症原代细胞培养及纤维化相关蛋白检测

doi:10.3969/j.issn.1006-7795.2019.04.020

首都医科大学学报 ›› 2019, Vol. 40 ›› Issue (4): 602-608.doi: 10.3969/j.issn.1006-7795.2019.04.020

子宫内膜异位症原代细胞培养及纤维化相关蛋白检测

张艳芹, 吴迪, 邓梦琪, 常翔宇, 苗劲蔚

首都医科大学附属北京妇产医院妇瘤科, 北京 100006

收稿日期:2019-02-20 出版日期:2019-07-21 发布日期:2019-07-19
通讯作者: 苗劲蔚 E-mail:miaojinweigyn@163.com
基金资助:
国家自然科学基金（81771549）。

Primary culture and detection of fibrosis associated proteins of endometriosis cells

Zhang Yanqin, Wu Di, Deng Mengqi, Chang Xiangyu, Miao Jinwei

Department of Gynecologic Oncology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100006, China

Received:2019-02-20 Online:2019-07-21 Published:2019-07-19
Supported by:
This study was supported by National Natural Science Foundation of China(81771549).

摘要/Abstract

摘要： 目的检测在位子宫内膜组织、子宫内膜异位症（endometriosis，EMs）异位病灶组织及正常人子宫内膜组织分离提纯子宫内膜间质细胞及相应间质细胞中纤维化相关蛋白表达水平及差异。方法在严格无菌条件下，刮勺刮取新鲜子宫内膜组织，冰盒运输，采用改良EMs原代细胞培养方法分离培养。并通过细胞爬片免疫组织化学方法，对分离提纯的细胞进行鉴定。蛋白免疫印迹方法检测对比分离提纯的细胞与相应组织纤维化相关蛋白表达的差异。结果 EMs在位内膜组织间质细胞原代分离12例均成功，成功率100%（成功指标：指细胞在培养瓶中贴壁生长并稳定传代）。EMs异位组织分离培养12例成功11例，成功率为91.7%。正常人子宫内膜组织分离9例成功8例，成功率为88.9%。人子宫内膜异位症异位间质细胞（human ectopic endometrial stromal cells，HEcESCs）与人子宫内膜异位症在位间质细胞（human eutopic endometrial stromal cells，HEuESCs）及正常人在位子宫内膜间质细胞（human normal endometrial stromal cells，HEnESCs）普通光镜观察无明显差异。但免疫组织化学、蛋白免疫印迹结果表明：HEcESCs纤维化相关蛋白的表达水平明显高于HEuESCs及HEnESCs两者纤维化相关蛋白表达水平（P<0.01），而HEuESCs与HEnESCs纤维化相关蛋白的表达差异无统计学意义（P>0.05），各间质细胞纤维化相关蛋白的表达与其相应组织水平相一致。结论采用改良EMs间质细胞原代培养方法，可以降低EMs子宫内膜间质细胞培养难度，并保持较高的细胞培养成功率。HEcESCs较HEuESCs及HEnESCs纤维化相关蛋白表达明显增高，即子宫内膜异位症患者，异位病灶在组织水平出现了明确的纤维化。

关键词: 子宫内膜异位症, 原代培养, 纤维化

Abstract: Objective To isolate and purify endometrial stromal cells from eutopic endometrium,ectopic tissue of patients with endometriosis (EMs) and normal endometrium. Detecting the expression of fibrosis-related protein in different tissues and corresponding stromal cells. Methods Fresh endometrial tissue was collected and modified EMs primary cell culture method was used. The cells were identified by immunohisto chemsistry. Western blotting was used to detect the consistency of fibrosis-related protein expression between the purified cells and the corresponding tissues. Results The primary isolation of stromal cells from endometrial tissue of EMs in situ was successful in 12 cases,the success rate was 100% (success index:cells adhered to the culture bottle and grew steadily). EMs ectopic tissue isolation and culture 12 successful 11 cases,the success rate of 91.7%. Eight cases of normal endometrial tissue were successfully isolated from 9 cases,with a success rate of 88.9%. There was no significant difference between human ectopic endometrial stromal cells (HEcESCs) and human eutopic endometrial stromal cells (HEuESCs) and normal human endometrial stromal cells (HEnESCs) under ordinary light microscope. But,Western blotting results showed that (P>0.05) the expression of fibrosis-related proteins in HEcESCs was significantly higher than that in HEuESCs and HEnESCs (P<0.01),and the expression of fibrosis-related proteins in stromal cells was consistent with there corresponding tissue levels. Conclusion The improved primary culture method of EMs cells can reduce the difficulty of EMs endometrial stromal cells culture,and maintain a high success rate of cell culture. The expression of fibrosis related protein in HEcESCs was significantly higher than that in HEuESCs and HEnESCs,that is mean,in patients with endometriosis,the ectopic lesions showed clear fibrosis at the tissue level.

Key words: endometriosis, primary culture, fibrosis

中图分类号:

R711.71

张艳芹, 吴迪, 邓梦琪, 常翔宇, 苗劲蔚. 子宫内膜异位症原代细胞培养及纤维化相关蛋白检测[J]. 首都医科大学学报, 2019, 40(4): 602-608.

Zhang Yanqin, Wu Di, Deng Mengqi, Chang Xiangyu, Miao Jinwei. Primary culture and detection of fibrosis associated proteins of endometriosis cells[J]. Journal of Capital Medical University, 2019, 40(4): 602-608.

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子宫内膜异位症原代细胞培养及纤维化相关蛋白检测

Primary culture and detection of fibrosis associated proteins of endometriosis cells

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