The therapeutic potential of umbilical cord mesenchymal stem cells in mice premature ovarian failure

Shufang Wang, Ling Yu, Min Sun, Sha Mu, Changyong Wang, Deqing Wang, Yuanqing Yao, Shufang Wang, Ling Yu, Min Sun, Sha Mu, Changyong Wang, Deqing Wang, Yuanqing Yao

Abstract

Mesenchymal stem cells, which are poorly immunogenic and have potent immunosuppressive activities, have emerged as promising cellular therapeutics for the treatment of several diseases. Mesenchymal-like cells derived from Wharton's Jelly, called umbilical cord matrix stem cells (UCMSCs), reportedly secrete a variety of cytokines and growth factors, acting as trophic suppliers. Here, we used UCMSCs to treat premature ovarian failure (POF). Ovarian function was evaluated by ovulation and the number of follicles. Apoptosis of the granulosa cells (GC) was analyzed by TUNEL staining. We found that after transplantation of the UCMSCs, apoptosis of cumulus cells in the ovarian damage model was reduced and the function of the ovary had been recovered. The sex hormone level was significantly elevated in mice treated with UCMSCs. The number of follicles in the treated group was higher than in the control group. Our results demonstrate that UCMSCs can effectively restore ovary functionality and reduce apoptosis of granulosa cells. We compared the RNA expression of the UCMSCs treated group with the POF model and wild-type control group and found that the UCMSC group is most similar to the wild-type group. Our experiments provide new information regarding the treatment of ovarian function failure.

Figures

Figure 1
Figure 1
Isolation and identification of UCMSCs. (a) The UCMSCs were isolated from human umbilical cord and showed typical fibroblastic morphology. (b) FCM analysis of UCMSCs. CD90, CD44, CD29, and CD105 were positive and CD31, HLA-DR were negative. Scale bars: 100 μm.
Figure 2
Figure 2
The effect of transplantation. (a, b, c) Hematoxylin eosin staining. (a) Follicles of the POF model. (b) Follicles of the WT group. (c) Follicles of the UCMSCs treated group. (d) Oocytes of the POF model. (e) Oocytes of the WT group. (f) Oocytes of the UCMSCs treated group. (g) Number of follicles of the three groups (POF, WT and UCMSCs treated). Data were means ± SD of numerous experiments. (POF versus WT P < 0.01, POF versus UCMSC P < 0.01, and UCMSC versus WT P > 0.05). (h) The values of sexual hormone E2 of the three groups (POF, WT and UCMSCs treated). Data were means ± SD of numerous experiments. (POF versus WT P < 0.05, POF versus UCMSC P < 0.05, and UCMSC versus WT P > 0.05). Scale bars: 100 μm.
Figure 3
Figure 3
Apoptosis of granulosa cells around follicles. (a, b, c) Green fluorescence indicates the apoptosis of granulosa cells in the three groups (POF, WT, and UCMSCs treated). (d, e, f) Blue fluorescence indicates the nucleus of granulosa cells. Scale bars: 50 μm. The green stained by POD kit. The blue stained by Hoechst.
Figure 4
Figure 4
RNA array analyses. (a) UCMSC group has 16 genes higher expression than WT group and 15 genes lower expression than WT group. Red: gene upregulated. Green: gene downregulated. (b) POF group has 33 genes higher expression than WT group and 58 genes lower expression than WT group. Red: gene upregulated. Green: gene down-regulated. (c) Genes relative expression of the three groups.
Figure 5
Figure 5
Genes correlation of three pathways. (a) UCMSC-WT gene Pathway Network. (b) POF-UCMSC gene Pathway Network. (c) POF-WT gene Pathway Network.

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