Isolation and Identification of Stromal Stem Cells from Eutopic Endometrium Functionional layer of Women with Endometriosis

Abstract

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. Study design: The endometrial samples from women with EMs undergoing fractional curettage respectively. Colony-forming cells (CFCs) were isolated from individual large colony-forming units (CFU) which visible macroscopically and contained >4000 cells. We focused on the difference of CFCs characteristics in women with EMs, including high proliferative potential（MTT assay）, serial clone formation, mammosphere formation of self-renewal, differentiation potential in vitro, and tissue reconstitution in vivo. Result: the optical densities in cell proliferation were more significant higher in the CFCs than N-CFCs ), The CE of CFCs from P1-5 are higher than N-CFCs 。There are statistical significances between the two groups (P<0.05). CFCs originated spheres with a diameter were at least 60μm, 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 did not change much in size after plating for 1 passage, and cannot be passaged serial .The protein 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. Conclusion: There are stem cells (SC) 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.

Key words: Endometriosis, Endometrial stem cells, Eutopic endometrial, Colony-forming assay

Ji-wen ZHANG. Isolation and Identification of Stromal Stem Cells from Eutopic Endometrium Functionional layer of Women with Endometriosis[J]. Journal of International Obstetrics and Gynecology, 2017, 44(1): 113-117.

References

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