Isolation, Cultivation Of Human Endometrial Stem/Progenitor Cells And Observation Of Xenotransplantation Human Endometrial Cells

Background: Repair and regeneration of the endometrium is related to endometrialstem cells (hESC), which maintain its function of menstruation and nurture a fertilizedegg. Some of the clinical disease may also be related to endometrial stem cells, suchas endometriosis (EMS). Some scholars have proposed EMS stem cell origin theorythat the blood back delivered the endometrial stem cells to places outside the uterinecavity, adhesion occurs in these seeds under suitable conditions, proliferation,invasion and growth of tumor formation. Scholars have been carried out to establishthe use of nude models in EMS, but because of its immune dysfunction, and lack ofrepresentation.Objective: The experiment will initially isolated human endometrial cells (hEC), andtransplanted the human endometrial cells into damaged seroua on the uterus of ICRmouse peritoneal,Are there lesionsformationwas observed after xenograft.Methods: Differential adhesion and quadratic sieve to isolated endometrial epithelialand stromal cell, then low-density respectively and picked clones to subculture,identified them by flow cytometry, immunohistochemistry. The isolated hECtransplant in serous damage zone of the uterus, contralateral uterus as a control.Experiment were divided into three groups: the first group transplanted ordinarysubcultured mixed hEC; The second group transplanted fresh separated hEC; Thethird group transplanted minced endometrial debris about0.5-1mm3size. In eachgroup, mice were sacrificed after2W,3W,4W. HE staining, immunohistochemicalidentification of anti-human ERα.To observe the effect of colonization and localreaction in the peritoneal cavity of the hESC.Results:①differential adhesion method and quadratic sieve filtration may initiallyseparated epithelial and stromal cells, epithelial cells are polygonal or oval and cellsarranged in cobblestone appearance after adhesion. Stromal cells are fusiform andcells arranged in spiral or radial. Cell respectively low density culture and both hadtwo size clones formed, and there are several different forms of cell clones. Largeclones were picked to subculture. epithelial cells began to senescent after P6,while stromal cells began to showed limited proliferation with in P9.Immunostaining wasfound P5lower cell CK-18expression, ABCG2, CD90and vimentin negative. Aftermixing the cells by flow cytometry subculture gradually reduce the proportion of SPcells was found to almost nothing.②After mixing subculture hEC transplant,2w,3w,4w mice were sacrificed showed no lesion formation, immunohistochemical analysiswas no anti-human ERα and CK expression. Freshly isolated hEC transplant digestedwith the same period last group of mice were sacrificed and the visible surface of theuterus have nodules, nodules were observed asanas were blister-like, HE staining ofblood vessels and interstitial tissue, no gland-like structures, immunohistochemistryno staining anti-human ERα and CK expression. Fresh endometrial tissue transplantmicro mice were sacrificed at3W visible at the surface of the uterus transplantnodules, nodules were observed asanas were blister-like surface bloodshot, HEstaining of blood vessels and interstitial tissue, no glands like structure.Immunohistochemical staining and CK regional anti-human ERα expression.Conclusion:1. human endometrial cells containing SP cells, these cells account forabout1.91percent, which are lost gradually in the process of subculture under normalculture medium and high oxygen environment.2.Human endometrial tissuetransplantation may be a slight short-term growth opportunity in ICR mice survivedthe womb.