Experimental Study On Transplantation Of Umbilical Cord Mesenchymal Stem Cells In The Treatment Of Premature Ovarian Failure

Premature ovarian failure (POF) is a syndrome signified by amenorrhea and genital atrophy before the age of 40 years, associated with low levels of gonadal hormones estrogens and high levels of the gonadotropins LH and FSH. POF is the common disease and frequently-occurring disease in obstetrics and gynecology clinical work, has become the main causes of infertility women today.Presently, hormone therapy and immune therapy are the main methods to restore ovarian function. But the treatment effect is temporary and transient, cannot effectively promote the repair damage of the ovarian tissue regeneration. For the limitations of conventional treatment methods, scientists are now actively looking for new and effective treatment. Mesenchymal stem cell therapy technology is an important branch of stem cell therapy technology, is considered to be the new hope of organ damage repair in the future.Here we used the HuMSCs single transplantation and multiple transplantation to therapy the POF, and preliminary discussion the possible mechanism. In this study,120mg/kg cyclophosphamide and 30mg/kg busulfan was administrated with the POF group mice by intraperitoneal injection. Anoestrum extended, the levels of estrogen decreased, the levels of the gonadotropins FSH increased, ovarian atrophy, the number of follicles reduced, the atretic follicle increased, AMH the secretion of granulosa cells reduced, the number of proliferating cells decreased in POF mice, which showed that successfully established a premature ovarian failure mice model. The umbilical cord were collected from mature infants. The specific surface markers of 5th HuMSCs passage cells were analyzed by flow cytometer. Cells positively expressed the specific surface markers CD44、CD73、 CD90 and CD105; and CD11b、CD19、CD34、CD45 and HLA-DR were negatively expressed. The HuMSCs differentiated into the osteoblasts and adipocytes. Successfully established HuMSCs lines. We used our HuMSCs lines single transplantation and multiple transplantation to therapy the POF. In single transplantation group, the sterilized mice had a single tail intravenous injection with 2 × 107/kg HuMSCs at 7 days post-induction. Each group mice were executed at 14,21,28,60 days post-induction. In single transplantation group, the sterilized mice had a single tail intravenous injection with 2x 107/kg HuMSCs at 7 days post-induction. Each group mice were executed at 14, 21,28,60 days post-induction.Compared with the POF group, organ coefficient of MSCs group only at 14 days post-induction obviously recovered, Primordial follicles, primary follicles, secondary follicles and mature follicles increased at 14,21,28,60 days post-induction, and the mature follicles dramaticlly increased at each time point.The levels of estrogen of MSCs group remarkably increased at 21,28,60 days post-induction., the levels of FSH dramaticlly decreased only at 60 days. AMH which represent ovarian reserve capacity observably increased in granulosa cells, Ki67 which represent cell proliferative activity significantly increased. Animal mating experiment results show that female fertility increased significantly. In multiple transplantation group, the sterilized mice were administrated tail intravenous injectionwith 2×107/HuMSCs three times at 7, 14,21 days post-induction. Each group mice were executed at 28,60 days post-induction. Compared with the POF group, organ coefficient of MSCs group obviously recovered at 28,60 days post-induction, Primordial follicles, primary follicles, secondary follicles and mature follicles increased at 14,21,28,60 days post-induction, and the Primordial follicles and primary follicles dramaticlly increased. The levels of estrogen and FSH of MSCs group remarkably recovered at 28,60 days post-induction., ovarian reserve capacity and ovarian cell proliferation significantly increased, female fertility recovered significantly. Mesenchymal stem cells were detected in follicular granulosa cells with tracer of the ovarian tissue freezing slice, which show HuMSCs to selectively migration and stay in damaged tissues. To sum up, HuMSCs transplantation can treat chemotherapy drugs cause POF, organ coefficient and hormone levels recovery effect in multiple transplantation mice were superior to single transplantation mice.