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Effect of Local Basic Fibroblast Growth Factor and Vascular Endothelial Growth Factor on Subcutaneously Allotransplanted Ovarian Tissue in Ovariectomized Mice.

Gao J, Huang Y, Li M, Zhao H, Zhao Y, Li R, Yan J, Yu Y, Qiao J - PLoS ONE (2015)

Bottom Line: The AC-3-positive in control group was significantly higher compared with bFGF group and VEGF group, and in bFGF+VEGF group was significantly lower than bFGF group and VEGF group.Survivin mRNA expression in bFGF+VEGF group was significantly higher than control group.The combination of bFGF and VEGF has beneficial effects on follicle survival, angiogenesis, and resumption of estrous cycles.

View Article: PubMed Central - PubMed

Affiliation: Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China; Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, 100191, China.

ABSTRACT

Objective: One of the major obstacles to ovarian tissue preservation is delayed angiogenesis that leads follicles lost after transplantation. The aim of the present study was to investigate the effects of bFGF and VEGF on heterotopic transplanted ovarian tissue using a mouse model.

Methods: Female mice underwent bilateral ovariectomy. Ovarian tissues encapsulated by fibrin hydrogels were transplanted subcutaneously into recipient mice, in which ovarian hormonal cyclicity was absent. The fibrinogen solution was mixed with bFGF, VEGF, or a mixture of bFGF and VEGF. The grafts were recovered 21 days after transplantation. Follicle morphology and follicle numbers were observed by H&E staining. Blood vessels were observed in transplanted intra-ovarian tissue by CD31 antibody IHC staining. Daily vaginal cytology was performed to determine estrous cycle and functional restoration of transplanted ovarian tissue. Blood was collected weekly and serum FSH levels were measured with a radioimmunoassay kit. Apoptosis analysis was performed by anti-AC-3 staining and survivin mRNA expression.

Results: The number of primordial follicles and secondary follicles in the bFGF+VEGF group was significantly higher than in the control group. The vascular density in the bFGF+VEGF groups were significantly higher than in the bFGF and the VEGF groups; there was no significant difference between the bFGF and VEGF groups. Estrous cycle was earlier in the bFGF+VEGF group compared with the control group; all mice in this group restored ovarian function. Serum FSH levels in the bFGF+VEGF group were significantly lower than in the control group by day 14 post-transplantation. The AC-3-positive in control group was significantly higher compared with bFGF group and VEGF group, and in bFGF+VEGF group was significantly lower than bFGF group and VEGF group. Survivin mRNA expression in bFGF+VEGF group was significantly higher than control group.

Conclusion: The combination of bFGF and VEGF has beneficial effects on follicle survival, angiogenesis, and resumption of estrous cycles.

No MeSH data available.


Ovarian tissue morphology after transplantation.(A) Representative micrograph of H&E-stained ovarian sections showing the preserved follicles. Original magnification100×. Scale bar = 100 μm. (B) The quantity of normal follicles at different developmental stages, including primordial, primary, and secondary follicles. * and # indicate that the p-value was <0.05 compared with the control group for primordial and secondary follicles, respectively.
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pone.0134035.g001: Ovarian tissue morphology after transplantation.(A) Representative micrograph of H&E-stained ovarian sections showing the preserved follicles. Original magnification100×. Scale bar = 100 μm. (B) The quantity of normal follicles at different developmental stages, including primordial, primary, and secondary follicles. * and # indicate that the p-value was <0.05 compared with the control group for primordial and secondary follicles, respectively.

Mentions: After 3 weeks, all grafts were retrieved, and most were found to be bigger than their original tissue size. There were even large antral follicles at the periphery of ovarian tissue grafts, indicating that the follicles had grown after transplantation and that ovulation would possibly occur soon (Fig 1A). The number of primordial and secondary follicles in the bFGF group and in the VEGF group was higher than that in the control group, but no significantly difference were found. The number of primordial and secondary follicles in the bFGF+VEGF group was significantly higher than in the control group, and was higher than in the bFGF and VEGF groups though there was no significant difference (Fig 1B). There were no differences among the four groups with regards to the number of primary follicles.


Effect of Local Basic Fibroblast Growth Factor and Vascular Endothelial Growth Factor on Subcutaneously Allotransplanted Ovarian Tissue in Ovariectomized Mice.

Gao J, Huang Y, Li M, Zhao H, Zhao Y, Li R, Yan J, Yu Y, Qiao J - PLoS ONE (2015)

Ovarian tissue morphology after transplantation.(A) Representative micrograph of H&E-stained ovarian sections showing the preserved follicles. Original magnification100×. Scale bar = 100 μm. (B) The quantity of normal follicles at different developmental stages, including primordial, primary, and secondary follicles. * and # indicate that the p-value was <0.05 compared with the control group for primordial and secondary follicles, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4514621&req=5

pone.0134035.g001: Ovarian tissue morphology after transplantation.(A) Representative micrograph of H&E-stained ovarian sections showing the preserved follicles. Original magnification100×. Scale bar = 100 μm. (B) The quantity of normal follicles at different developmental stages, including primordial, primary, and secondary follicles. * and # indicate that the p-value was <0.05 compared with the control group for primordial and secondary follicles, respectively.
Mentions: After 3 weeks, all grafts were retrieved, and most were found to be bigger than their original tissue size. There were even large antral follicles at the periphery of ovarian tissue grafts, indicating that the follicles had grown after transplantation and that ovulation would possibly occur soon (Fig 1A). The number of primordial and secondary follicles in the bFGF group and in the VEGF group was higher than that in the control group, but no significantly difference were found. The number of primordial and secondary follicles in the bFGF+VEGF group was significantly higher than in the control group, and was higher than in the bFGF and VEGF groups though there was no significant difference (Fig 1B). There were no differences among the four groups with regards to the number of primary follicles.

Bottom Line: The AC-3-positive in control group was significantly higher compared with bFGF group and VEGF group, and in bFGF+VEGF group was significantly lower than bFGF group and VEGF group.Survivin mRNA expression in bFGF+VEGF group was significantly higher than control group.The combination of bFGF and VEGF has beneficial effects on follicle survival, angiogenesis, and resumption of estrous cycles.

View Article: PubMed Central - PubMed

Affiliation: Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China; Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, 100191, China.

ABSTRACT

Objective: One of the major obstacles to ovarian tissue preservation is delayed angiogenesis that leads follicles lost after transplantation. The aim of the present study was to investigate the effects of bFGF and VEGF on heterotopic transplanted ovarian tissue using a mouse model.

Methods: Female mice underwent bilateral ovariectomy. Ovarian tissues encapsulated by fibrin hydrogels were transplanted subcutaneously into recipient mice, in which ovarian hormonal cyclicity was absent. The fibrinogen solution was mixed with bFGF, VEGF, or a mixture of bFGF and VEGF. The grafts were recovered 21 days after transplantation. Follicle morphology and follicle numbers were observed by H&E staining. Blood vessels were observed in transplanted intra-ovarian tissue by CD31 antibody IHC staining. Daily vaginal cytology was performed to determine estrous cycle and functional restoration of transplanted ovarian tissue. Blood was collected weekly and serum FSH levels were measured with a radioimmunoassay kit. Apoptosis analysis was performed by anti-AC-3 staining and survivin mRNA expression.

Results: The number of primordial follicles and secondary follicles in the bFGF+VEGF group was significantly higher than in the control group. The vascular density in the bFGF+VEGF groups were significantly higher than in the bFGF and the VEGF groups; there was no significant difference between the bFGF and VEGF groups. Estrous cycle was earlier in the bFGF+VEGF group compared with the control group; all mice in this group restored ovarian function. Serum FSH levels in the bFGF+VEGF group were significantly lower than in the control group by day 14 post-transplantation. The AC-3-positive in control group was significantly higher compared with bFGF group and VEGF group, and in bFGF+VEGF group was significantly lower than bFGF group and VEGF group. Survivin mRNA expression in bFGF+VEGF group was significantly higher than control group.

Conclusion: The combination of bFGF and VEGF has beneficial effects on follicle survival, angiogenesis, and resumption of estrous cycles.

No MeSH data available.