国际妇产科学杂志2017，Vol.44Issue(1)：113-117,5.
人子宫内膜异位症在位子宫内膜干细胞的分离和鉴定
Isolation and Identification of Stromal Stem Cells from Eutopic Endometrium Functionional Layer of Women with ;Endometriosis
摘要
Abstract
Objective: To get the endometrium stem cells (ESC) for the eutopic endometrial from women with endometriosis (EMs) by using colony-forming assay and provide the experimental basis for the further study of EMs. Methods:The endometrial samples from women with EMs who treated in Department of Obstetrics and Gynecology, Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine in 2012-2014 undergoing fractional curettage respectively. Colony-forming cells (CFCs) were isolated from individual colony-forming units (CFU) which visible macroscopically and contained >4 000 cells. We focused on the characteristics of stem cells between CFCs and N-CFCs, including high proliferative potential (MTT assay), serial clone formation, mammosphere formation of self-renewal, differentiation potential in vitro, and tissue reconstitution in vivo. Results:The optical densities in cell proliferation were more significantly higher in the CFCs than N-CFCs (P<0.05). The cloning efficiency (CE) of CFCs from P1-5 are higher than N-CFCs [CFCs:(12.30±3.75)%, (13.11±3.23)%, (11.86±2.21)%, (12.08±3.01)%, (11.01±3.98)%; N-CFCs:(5.57±2.13)%, (4.21±2.12)%, (2.23±1.02)%, (0.12 ±0.07)%, 0%]. There are statistical significances between the two groups (P<0.05). CFCs originated spheres with a diameter were at least 60 m, spheres were continuously generated at 5-6 day intervals for at least 3 passages with a bigger and bigger size,in contrast, the spheres of N-CFCs cannot be passaged serial [Count the cells of each passage:CFCs:(0.82±0.37)× 104, (1.45±0.64)×104, (3.06±1.73)×104, N-CFCs:(0.48±0.27)×104, (0.37±0.15)×104, 0]. There are statistical significances between the two groups (P<0.05). The protein expression of CFCs was negative for CK-19 before induction culture, but were expressed at CFCs, which cultured with the induction medium for 21 days. The endometriotic lesion-forming efficiency (100%) in the CFCs group was higher than that of N-CFCs group (40.63%), when CFCs and N-CFCs were inoculated into the abdominal subcutaneous of nude mice, respectively. Reconstruction of endometriotic lesions in the CFCs group with an average volume at (30.95 ±12.13)mm3 was bigger than the N-CFCs group (19.12±10.98)mm3. Conclusions:There are stem cells in the function layer of endometrium in EMs patients and it can be isolated from eutopic endometrium using colony-forming assays. The study has given a simple method to get ESC easily for researching EMs. Endometrial stem cells can be cryopreserved and subcultured, and have the ability of unlimited proliferation, self-renewal, directional differentiation and ectopic lesion fomation.关键词
子宫内膜异位症/子宫内膜/干细胞/集落形成单位测定Key words
Endometriosis/Endometrium/Stem cells/Colony-forming units assay引用本文复制引用
张继雯,宋殿荣,张崴,刘三洪,王玉华,杜文欣,郭洁,王雅楠..人子宫内膜异位症在位子宫内膜干细胞的分离和鉴定[J].国际妇产科学杂志,2017,44(1):113-117,5.基金项目
中国博士后科学基金（201104309）；国家自然科学基金 （）
