Preliminary Investigation Of Human Umbilical Cord Mesenchymal Stem Cells In Premature Ovarian Failure Therapy

Premature ovarian failure (POF) is a syndrome signified by amenorrhea beforethe age of40years. POF is generally characterized by menstrual disorders, loss oflibido, decreased sexual function, infertility, low levels of gonadal hormonesestrogens and inhibins and high levels of the gonadotropins LH and FSH,hypergonadotropic amenorrhea. Representing one major cause of female infertility.Here we used the umbilical cord mesenchymal stem cells (UCMSCs) and theconditioned medium (CM) of UCMSCs to therapy the POF. This article includes fourparts. The first part is the research of UCMSCs in the treatment of POF. The secondpart is the research of CM in the treatment of POF. The third part is the research ofUCMSCs and CM in the treatment of POF, especially in the improvement of oocyte.The fourth part is to investgate the mechanism of UCMSCs and CM in POF therapy.Part1Research of UCMSCs in the treatment of chemotherapy POF mousemodalObjective: The aim of this study was to objectively evaluate the benefits ofUCMSCs for the premature ovarian failure in mice model. Method: Wild-typefemale mice with ICR background (6-8weeks old) were randomly into groups A, B, Cand D (n=20)(A, B and C were treated with CTX to induce the chemotherapy POFmouse modal, while those in group A received no treatment. The USMSCs weretransplanted into the mice of group B and C, B was injected via the ovarian situ, Cwas injected via the tail vein. D was the control group.) Comparison the variousindexes differences of four groups, these indexes include: the number of follicles, HEstaining of ovarian tissue and number of apoptotic cells in the ovarian tissue. Results:Successfully established UCMSCs lines and a cyclophosphamide-induced prematureovarian failure in mice model; After treatment, UCMSCs could promote the repair ofthe ovarian injury, the number of follicles and quality have been significantly improved, at the same time, the number of apoptotic cells was significantly reduced inthe injured ovaries. Conclusion: Our study shows that UCMSCs have obvioustherapeutic effect for the POF mice model; they Can effectively improve the ovarianfunction by increased the number of follicles, improved the quality of follicles, andreduced the number of apoptotic cells.Part2Research of CM in the treatment of chemotherapy POF mouse modalObjective: To investigate the application of CM in the treatment of POF.Methods: Select the most vigorous growth UCMSCs, the medium was replaced withserum-free medium. Continued to culture the cells for48hours, then collected theCM. Wild-type female mice with ICR background (6-8weeks old) were randomly intofour groups A, B, C and D (n=20)(A, B and C were treated with CTX to induce thechemotherapy POF mouse modal, while those in group A received no treatment. TheCM was injected into the mice of group B via the tail vein, mice in group C wasinjected MSCGD via the tail vein. The equal volume of PBS was injected by the tailvein in the mice of group A and D.) Comparison the various indexes differences offour groups, in order to observe the treatment of UCMSCs for the POF modal, theseindexes include: the number of follicles, HE staining of ovarian tissue and number ofapoptotic cells in the ovarian tissue. Results: Successfully obtained the CM from theUCMSCs; after treatment, CM could promote the repair of the ovarian injury, thenumber of follicles and quality have been significantly improved, at the same time,the number of apoptotic cells was significantly reduced in the injured ovaries.Conclusions: Our study shows that CM has obvious therapeutic effect for the POFmice model, but MSCGD did not play a therapeutic role. So, there should be somematerial in CM secreted from UCMSCs which can effectively improve the ovarianfunction by increased the number of follicles, improved the quality of follicles, andreduced the number of apoptotic cells.Part3Effect of UCMSCs and CM transplantation on immunological injury ofthe ovary in mice.Objective: To investigate the effect of UCMSCs and CM in repairing ovarianinjury in mice sensitized with the peptides of the mouse zona pellucid3(ZP3). Methods: Wild-type female mice with ICR background (6-8weeks old) wererandomly into six groups A, B, C, D, E and F (n=20)(A, C, D and E were treatedwith ZP3and CFA to induce the immunological POF mouse modal, while those ingroup A received no treatment. The CM was injected into the mice of group C via thetail vein, mice in group D was injected with MSCGD via the tail vein, UCMSCs wasinjected into the mice of group E also via the tail vein. Mice in group B were Injectedsubcutaneously only with CFA. The equal volume of PBS was injected by the tailvein in the mice of group A, B and F, F was the control group.) Result: To observethe number and quality of follicle, the number and quality of oocytes, we found thatUCMSCs concentrate on the treatment of premature ovarian failure. Conclusions:UCMSCs and CM can improve the immunological injury of the ovary in mice,especially in the number and quality of oocytes. That will be a new approach for thetreatment of premature ovarian failure patients, to better restore the patient's fertility.Part4to investgate the mechanism of UCMSCs and CM in POF therapyObjective: To objectively evaluate the benefits of UCMSCs for the prematureovarian failure in chemotherapy mouse model, in order to reveal the specificmechanisms to improve ovarian function. Methods: Wild-type female mice with ICRbackground (6-8weeks old) were randomly into three groups A, B and C (n=20)(Aand B were treated with CTX to induce the chemotherapy POF mouse modal, whilethose in group A received no treatment. The UCMSCs was injected into the mice ofgroup B via the tail vein, the equal volume of PBS was injected by the tail vein in themice of group A and C, C was the control group.) Result: Mouse Genome Arrayscreened a number of differentially expressed genes, some of them play importantrole in the metabolic pathway of life. Conclusions: UCMSCs can change the level ofexpression of certain genes in the ovarian tissue, thus affecting ovarian function, andlaid the foundation for us to further explore the mechanism of UCMScs treatment ofpremature ovarian failure.