The Effects of Human Menstrual-Derived Stem Cells Transplantation on Premature Ovarian Failure in Rats

Background: Premature ovarian failure (POF) is one of the problems that young women faced during chemotherapy. POF is a common disorder caused by depletion of the ovarian re-serve, which results in menopause and infertility. Among the various treatment methods, the use of mesenchymal stem cells is recognized as a suitable treatment. Mesenchymal stem cells have therapeutic potential and can be derived from several sources, including bone marrow, adipose tissue, amniotic fluid and etc. All of them show the potential for restoring ovarian function and save long-term fertility in female mice treated with chemotherapy. But the method of collecting these cells is highly invasive so it is difficult to use in the clinic. Recently, the population of highly proliferative stem cells in menstrual blood were detected with similar properties of mesenchymal stem cells and are expressed without restrictions.Various stud-ies have been done on the therapeutic function of HuMenSCs in various diseases. This study examine the effect of menstrual blood stem cells on the improvement of function and ovarian regeneration in POF models.Materials and Methods: Female Rats were injected intraperi-toneally with 36 mg/kg busulfan. HuMenSCs were obtained, grown and analyzed for immunophenotypic features at pas-sage three. The cells were labelled with CM-Dil and infused into the rats, 7 days after i.p. injection of busulfan for homing assay. There were four groups: normal, negative control (the rats were administered busulfan), treatment (rats were injected intravenously with 1 × 106 autologous HuMenSCs in a volume of 1mm PBS) and Sham, (rats were injected intravenously with 1mm PBS via the tail vein). One month after treatment, the ovaries were collected and weighed. Histological sections were prepared from the ovary and HuMenSCs were tracking. Then the follicles were counted and classified. Ovarian function was evaluated by monitoring ovulation. Also Histological sections were stained for TUNEL test. Subsequently, we examined the changes of expression of BAX and BCL2 apoptotic genes by Real-time PCR assay.Results: One week after injection of busulfan, the ovaries were atrophied and evacuated from the follicle. Cultured HuMenSCs indicated a high level of expression of CD44 and CD90 but low levels of CD34 and CD45 (for all P≤0.05). One month after Hu-MenSCs transplantation, these cells were located in the ovar-ian interstitium and Granulosa cells and the number of follicles and ovary weight increased. Apoptosis was evaluated by Tun-nel staining and the expression level of Bax and BCL2 genes. The number of TUNEL-positive cells significantly decreased in treatment group (P<0.0001). And also the expression level of Bax genes significantly decreased compared to negative and sham groups (P<0.0001). There was also an increase in the lev-el of BCL2 gene expression in the treatment group.Conclusion: HuMenSCs improve and restore ovarian func-tion and reduce apoptosis in damaged ovarian tissue caused by busulfan toxicity. Since access to these cells is easy and non-invasive, therefore the use of these cells can be a practical and low-cost method for the treatment of POF patients.

Keywords:

POF , Humenscs , Busulfan , Apoptosis , Granulosa Cells