Primary culture and detection of fibrosis associated proteins of endometriosis cells

doi:10.3969/j.issn.1006-7795.2019.04.020

Abstract

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

CLC Number:

R711.71

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.

References 12

[1]	Grande G,Vincenzoni F, Milardi D, et al. Cervical mucus proteome in endometriosis[J]. Clin Proteom,2017,14(1):7.
[2]	Mishra V V,Gaddagi R A,Aggarwal R,et al. Prevalence; characteristics and nanagement of endometriosis amongst infertile women:a one year retrospective study[J]. J Clin Diagnost Res,2015,9(6):QC013.
[3]	中华医学会妇产科学分会子宫内膜异位症协作组. 子宫内膜异位症的诊治指南[J]. 中华妇产科杂志,2015,50(3):161-169.
[4]	Bulun S E. Endometriosis[J]. N Engl J Med,2009,360(3):268-279.
[5]	Singh S S,Suen M W. Surgery for endometriosis:beyond medical therapies[J]. Fertil Steril,2017,107(3):549-554.
[6]	Ziegler D D,Borghese B,Chapron C. Endometriosis and infertility:pathophysiology and management[J]. Lancet,2010,376(9742):730-738.
[7]	Brosens J J. Progesterone receptor regulates decidual prolactin expression in differentiating human endometrial stromal cells[J]. Endocrinology,1999,140(10):4809-4820.
[8]	俞超芹,石书芳,刘玉环,等. 人子宫内膜异位症在位和异位内膜细胞原代培养及形态学观察[J]. 中西医结合学报,2006,4(2):189-193.
[9]	Matthews C J, Redfern C P,Hirst B H,et al. Characterization of human purified epithelial and stromal cells from endometrium and endometriosis in tissue culture[J]. Fertil Steril,1992,57(5):990-997.
[10]	Koninckx P R,Barlow D,Kennedy S. Implantation versus infiltration:the Sampson versus the endometriotic disease theory[J]. Gynecol Obstetr Invest,1999,47(Suppl 1):3-10.
[11]	Dogan S,Machicao F,Wallwiener D,et al. Association of peroxisome proliferator-activated receptor gamma 2 Pro-12-Ala polymorphism with endometriosis[J]. Fertil Steril,2004,81(5):1411-1413.
[12]	张真真, 戴姝艳.血管紧张素Ⅱ2型受体对大鼠子宫内膜异位症血管生成的影响[J].中国医科大学学报,2018,47(9):792-796.

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