| Objective: The aim of is study was to explore the therapeutic effect of menstrual blood mesenchymal stem cells on thin endometrium after induced decidualization,providing a new idea for the treatment of thin endometrium.Methods:1.Collected female menstrual blood aseptically,Menstrual blood stem cells(MenSCs)were isolated,cultured,identified,and induced differentiation(mainly osteogenic,lipogenic,and chondrogenic).All the experiments were performed to determine the characteristics of these mesenchymal stem cells characterization.2.The MenSCs was induced decidualization in vitro and drawn two cell growth curves.After induced decidualization,q-PCR was used to detect the expression of decidualrelated genes(PRL and IGFBP-1)and endometrial receptivity related genes(LIF and HOXA10).The concentration of VEGF-A in the supernatant of the two cells was detected by ELISA.Finally,the function of VEGF-A was verified by scratch test,and the differences between different groups were counted.3.TE rat model was based on the traditional method,and the feasibility of the new method was determined by statistical analysis of endometrial thickness,number of glands and embryos in rats.4.MenSCs(n=3)and decidualized MenSCs(n=3)were transplanted into TE rat models established by the new method to treat TE model rats.Two weeks later,endometrial RNA was extracted to detect the expression of genes related to angiogenesis(VEGF-A)and cell proliferation(SCF、IGF-1).The endometrial thickness and the number of glands in different treatment groups were counted.Finally,MenSCs(n=15)and decidualized MenSCs(n=15)were transplanted into the TE rats endometrium,and the effect of treatment was reflected by the number of embryos.Evaluated the effect of decidualized MenSCs on TE rats’ fertility comprehensively.Results:1.MenSCs appear as spindle and clonal growth under the microscope observation.The flow cytometry showed that the MenSCs were positive for CD44,CD73,CD90,CD105 and negative for CD11 b,CD34,CD79 a,and the cells were successfully induced to differentiate into adipocyte,osteocyte and chondrocyte in vitro.Those results prove that the MenSCs own mesenchymal stem cell properties.2.PRL and IGFBP-1 genes were increased after induced decidualization(P ≤ 0.0001),LIF and HOXA10 were also increased(P ≤ 0.05),and the cell proliferation curve showed that the proliferation ability of cells after decidualization decreased compared with that of cells without decidualization.There was a higher concentration of VEGF-A in the supernatant of MenSCs and the decidualized cells than in the normal medium,and the VEGF-A secreted by decidual cells was higher than that of MenSCs(P ≤ 0.001).The function of VEGF-A was verified by scratch experiment.The results indicated that cell proliferation decreased but secretion increased after decidualization.3.The new TE method could lead to thinner endometrium,fewer glands and fewer embryos.4.Both decidualized cells and MenSCs can promote the formation of endometrial blood vessels,regenerate endometrial thickness and the number of glands in rats,but there is no significant difference in the statistical results(P > 0.05).MenSCs and decidualized cells both have a significant effect on improving the pregnancy rate of rats(P≤0.05),but the effect of decidualized cells was better than that of MenSCs.Conclusion:1.Decidualized MenSCs secreted more VEGF-A than MSC,and promote endometrium receptivity related genes expression.2.Both decidualized MenSCs and undecidualized MenSCs promote endometrial tissue proliferation,improve the number of glands and pregnancy rate in TE rats,and the mechanism may be related to the promotion of endometrial angiogenesis and cell proliferation.3.Decidualized MenSCs were superior to undecidualized MenSCs in improving uterine receptivity of rats,which provided a new idea for the clinical application of MenSCs in the treatment of fertility in TE patients. |
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