Bone marrow transplantation restores follicular maturation and steroid hormones production in a mouse model for primary ovarian failure

Mohsen Ghadami, Ebtehal El-Demerdash, Dong Zhang, Salama A Salama, Awadh A Binhazim, Anthony E Archibong, Xinlei Chen, Billy R Ballard, M Ram Sairam, Ayman Al-Hendy, Mohsen Ghadami, Ebtehal El-Demerdash, Dong Zhang, Salama A Salama, Awadh A Binhazim, Anthony E Archibong, Xinlei Chen, Billy R Ballard, M Ram Sairam, Ayman Al-Hendy

Abstract

Recent studies suggest that bone marrow stem cells (BMSCs) are promising grafts to treat a variety of diseases, including reproductive dysfunction. Primary ovarian failure is characterized by amenorrhea and infertility in a normal karyotype female, with an elevated serum level of follicle-stimulating hormone (FSH) and a decrease level of estrogen caused by a mutation in FSH receptor (FSHR) gene. Currently, there is no effective treatment for this condition. The phenotype of FSHR (-/-) mouse, FORKO (follitropin receptor knockout), is a suitable model to study ovarian failure in humans. Female FORKO mice have elevated FSH, decreased estrogen levels, are sterile because of the absence of folliculogenesis, and display thin uteri and small nonfunctional ovaries. In this study, we determined the effects of BMSC transplantation on reproductive physiology in this animal model. Twenty four hours post BMSC transplantation, treated animals showed detectable estroidogeneic changes in daily vaginal smear. Significant increase in total body weight and reproductive organs was observed in treated animals. Hemotoxylin and eosin (H&E) evaluation of the ovaries demonstrated significant increase in both the maturation and the total number of the follicles in treated animals. The FSH dropped to 40-50% and estrogen increased 4-5.5 times in the serum of treated animals compared to controls. The FSHR mRNA was detected in the ovaries of treated animals. Our results show that intravenously injected BMSCs were able to reach the ovaries of FORKO mice, differentiate and express FHSR gene, make FSHR responsive to FSH, resume estrogen hormone production, and restore folliculogenesis.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Amplified DNA from different tissues…
Figure 1. Amplified DNA from different tissues of animals transplanted by (+/+) or (−/−) donors.
Figure 2. Changes of total body weight(A),…
Figure 2. Changes of total body weight(A), ovaries(B), uterus(C), vagina and cervix(D), and serum level of FSH(E) and estrogen(F) in treated (Tr) Vs control (Ct) animals at different time points of experiment.
For B, C and D organs weight considered as % of total body weight. Both treated and control group had increase in total body weight but BMT group showed significantly more increase than control group (A, P

Figure 3. Development of the ovary in…

Figure 3. Development of the ovary in BMT animal (A) compared to untreated control animal…

Figure 3. Development of the ovary in BMT animal (A) compared to untreated control animal (B).
Both the total number of follicles and the number of antral follicles are significantly higher in BMT compare to control group. Histological evaluation showed on average 28±4 follicles/ovary in treated group with 8±2 follicles at the antral stage compared to only 6±2 with no follicles at antral stage in untreated control mice. Photos have been taken at the same magnification.

Figure 4. mRNA expression of FSHR gene…

Figure 4. mRNA expression of FSHR gene in the ovaries of treated and control animals…

Figure 4. mRNA expression of FSHR gene in the ovaries of treated and control animals and in non-reproductive organs (liver) at two different time points (1 and 12 weeks).
As shown, FSHR mRNA is only expressed in the ovaries of BM treated animals and there is no mRNA expression in the ovary of untreated and in non-reproductive organs of treated group.
Figure 3. Development of the ovary in…
Figure 3. Development of the ovary in BMT animal (A) compared to untreated control animal (B).
Both the total number of follicles and the number of antral follicles are significantly higher in BMT compare to control group. Histological evaluation showed on average 28±4 follicles/ovary in treated group with 8±2 follicles at the antral stage compared to only 6±2 with no follicles at antral stage in untreated control mice. Photos have been taken at the same magnification.
Figure 4. mRNA expression of FSHR gene…
Figure 4. mRNA expression of FSHR gene in the ovaries of treated and control animals and in non-reproductive organs (liver) at two different time points (1 and 12 weeks).
As shown, FSHR mRNA is only expressed in the ovaries of BM treated animals and there is no mRNA expression in the ovary of untreated and in non-reproductive organs of treated group.

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