子宫腺肌病原代内膜间质细胞培养及功能学鉴定

Culture and functional identification of primary mesenchymal cells in adenomyosis

目的分离提纯培养子宫腺肌病(AM)内膜间质细胞(ESc),检测其细胞功能的差异,为进一步的细胞及分子水平研究奠定基础。方法选取2013年1月至2016年12月份在本院治疗的正常子宫内膜和子宫腺肌病手术病例,在严格无菌条件下,收集临床确诊的AM患者在位子宫内膜组织12例、异位子宫内膜组织41例及正常对照内膜组织13例,分别分离培养ESc,并通过免疫组织化学法对分离提纯培养的细胞进行鉴定。分别检测三种细胞的增殖、迁移、侵袭和凋亡等细胞功能。结果正常人ESc 13例,AM患者在位12例、异位ESc原代分离41例均成功,成功率为100.00%。与正常在位ESc相比,在位、异位ESc在普通光镜下,形态观察无明显差异,但是细胞功能方面,在位ESc生长曲线显示增殖率明显升高;异位ESc的迁移、侵袭率水平升高,差异有统计学意义(P<0.01),三种细胞凋亡无明显差异。结论采用间质细胞原代培养方法可以成功培养AM ESc,并保持较高的细胞培养成功率。在位ESc增殖率最高,异位ESc的迁移、侵袭率水平升高,细胞凋亡均无明显差异。

Objective To isolate,purify and culture endometrial stromal cells(ESc)of adenomyosis(AM),and to detect the difference of cell function,so as to lay a foundation for further research at the cellular and molecular levels.Methods The normal endometrial and adenomyosis surgical cases treated in our hospital from January 2013 to December 2016 were selected.Under strict aseptic conditions,12 cases of eutopic endometrial tissues,41 cases of ectopic endometrial tissues and 13 cases of normal control endometrial tissues of clinically diagnosed patients with AM were collected.ESc were isolated and cultured,and the isolated and purified cells were identified by immunohistochemistry.The proliferation,migration,invasion and apoptosis of the three types of cells were detected.Results 13 cases of normal human ESc,12 cases of patients with eutopic AM and 41 cases of ectopic ESc were successfully isolated,with a success rate of 100.00%.Compared with normal eutopic ESc,the morphology of eutopic and ectopic ESc was not significantly different under ordinary optical microscope,but the proliferation rate of eutopic ESc was significantly increased in terms of cell function.The migration and invasion rates of ectopic ESc were significantly increased(P<0.01),and there was no significant difference among the three types of apoptosis.Conclusion AM ESc can be successfully cultured by using the primary culture method of mesenchymal cells,and a high success rate of cell culture can be maintained.The proliferation rate of eutopic ESc is the highest,the migration and invasion rate of ectopic ESc increases,and there is no significant difference in apoptosis.