Study Of The Repair Of Injuried Mouse Endometrium By Using Embryonic Stem Cells

Background: The difficulty in endomentrial repair is related to many gynecologydiseases, the clinical treatment of these diseases is very tough. Over the recent years,some scholars have put forward the hypothesis that the physiogenic regeneration andrepair of endomentrial is caused by the stem cells which indwell the endometrial zonabasalis.We can make further assumption that the fail of endomentrial repair is due tothe exhaustion or dysfunction of the stem cells in the basal layer. Therefore, weassume that we could help the repair of endomentrial by adding exogenous embodystem cells(ESC).Objective: Our experiment is to find out whether isotype variant embody stem cellsthat transplanted into acute injuried uterine will contribute to the repair ofendomentrum, how the ESC will affect the repair procedure of endomentrail, whetherthe ESC will colonize in the endomentrium and how long they will exsit, whether theESC will be induced to differentiate in the repair procedure, and whether the ESC willturn to tumor.Methods: We establish the modle of the mouse’s injured endometrium. Recoverythe eGFP-gene marked mouse embody stem cells, then we used the mouse embryonicfibroblast cells to feed it, after that we transplanted it into the kidney capsule anddetected its all-round identification. In general, the experiment was divided into threegroups. In the first group, the ESC was transplanted to both the damaged and theundamaged uterine of the mouse. In the second group, the ESC was injected to themouse that had only one damaged uterine by tail vein. In the third group, the ESC wascocultured with endometrial cells，and then they were transplanted to one uterine ofthe mice whose both uterine were damaged, simultaneity equivalent PBS wastransplanted into the other uterine. The mice were sacrificed at the1th week,2th week, 3th week,4th week respectively after the transplantation, then we observed thesurvival and migration status of the transplanted mouse embryonic stem cells directlyby fluorescence microscopy, and under the microscope we did HE staining andAnti-GFP antibody immunohistochmical staining.Result: The ESC grew well on the MEF feeder cells as clone-like, the eGFP-genemarked ESC was observed expressing high green fluorescence under fluorescencemicroscope and having teratoma formation with three germ layers by HE andimmunohistochmical staining. ESC could be differentia to thin and flat clone-likeendometrium epithelioid cells and observed with high green fluorescence underfluorescence microscope after cocultured with endometrial cells. We sacrificed themice after ESC was transplanted at a series of time, then high green fluorescence inthe damaged uterine was observed under the fluorescence microscope, and theundamaged uterine didn’t show fluorescence, but it developed teratoma and expressedgreen fluorescence as time went on. The mouse abdominal wall cut and damageduterine were observed green fluorescence under the fluorescence microscope afterESC was injected into the tail vein, no tumor appeared as time went on. The mice,that had been transplanted into ESC which was cocultured with endometrial cells,were observed green fluorescence in the cells transplanted uterine under thefluorescence microscope, and no tumor growth appeared during the observationperiod. The tissues were analyzed by the HE staining and Anti-GFP antibodyimmunohistochmical staining. In the first group, the tumor conclude fat, cartilage,gland, nervus like histodifferentiation. The second group, Anti-GFP antibodyimmunohistochmical staining positive cells could be found in the endomentrum likenest distribution in the uterine of the mice whose tail vein was injected ESC*. Thethird group, the damaged uterine which was transplanted into mixture cells repairedwell, and integrated endometrial epithelium and gland which Anti-GFP antibodyimmunohistochmical staining positive could be found in the endomentrum.Conclusions: ESC can survive in the damaged uterine cavity and can developteratoma. Injury is the most important factor in inducing embody stem cellsrecruitment. After the ESC was cocultured with endometrial cells and then transplanted to the damaged uterine, the mixed cells can survive in the damageduterine and the tumorigenicity was reduced.