RESEARCH ARTICLE
Effect of Local Basic Fibroblast Growth Factor and Vascular Endothelial Growth Factor on Subcutaneously Allotransplanted Ovarian Tissue in Ovariectomized Mice Jiangman Gao1,2, Ying Huang1,2, Min Li1, Hongcui Zhao1,3, Yue Zhao1,2, Rong Li1,2,3, Jie Yan1,3*, Yang Yu1,2*, Jie Qiao1,2,3 1 Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China, 2 Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, 100191, China, 3 Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Beijing, 100191, China * [email protected] (YY); [email protected] (JY)
Abstract OPEN ACCESS Citation: Gao J, Huang Y, Li M, Zhao H, Zhao Y, Li R, et al. (2015) Effect of Local Basic Fibroblast Growth Factor and Vascular Endothelial Growth Factor on Subcutaneously Allotransplanted Ovarian Tissue in Ovariectomized Mice. PLoS ONE 10(7): e0134035. doi:10.1371/journal.pone.0134035 Editor: Meijia Zhang, China Agricultural University, CHINA Received: March 19, 2015 Accepted: July 5, 2015 Published: July 24, 2015 Copyright: © 2015 Gao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper. Funding: This work was supported in part by the Ministry of Science and Technology of China Grants (973 program; 2011CB944504 and 2014CB943203), and the National Natural Science Funds for general program (31371521 and 31230047). Competing Interests: The authors have declared that no competing interests exist.
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
PLOS ONE | DOI:10.1371/journal.pone.0134035 July 24, 2015
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Growth Factor and Ovarian Tissue Transplantation
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.
Introduction In women, approximately one tenth of cancers occur in
Effect of Local Basic Fibroblast Growth Factor and Vascular Endothelial Growth Factor on Subcutaneously Allotransplanted Ovarian Tissue in Ovariectomized Mice Jiangman Gao1,2, Ying Huang1,2, Min Li1, Hongcui Zhao1,3, Yue Zhao1,2, Rong Li1,2,3, Jie Yan1,3*, Yang Yu1,2*, Jie Qiao1,2,3 1 Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China, 2 Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, 100191, China, 3 Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Beijing, 100191, China * [email protected] (YY); [email protected] (JY)
Abstract OPEN ACCESS Citation: Gao J, Huang Y, Li M, Zhao H, Zhao Y, Li R, et al. (2015) Effect of Local Basic Fibroblast Growth Factor and Vascular Endothelial Growth Factor on Subcutaneously Allotransplanted Ovarian Tissue in Ovariectomized Mice. PLoS ONE 10(7): e0134035. doi:10.1371/journal.pone.0134035 Editor: Meijia Zhang, China Agricultural University, CHINA Received: March 19, 2015 Accepted: July 5, 2015 Published: July 24, 2015 Copyright: © 2015 Gao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper. Funding: This work was supported in part by the Ministry of Science and Technology of China Grants (973 program; 2011CB944504 and 2014CB943203), and the National Natural Science Funds for general program (31371521 and 31230047). Competing Interests: The authors have declared that no competing interests exist.
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
PLOS ONE | DOI:10.1371/journal.pone.0134035 July 24, 2015
1 / 11
Growth Factor and Ovarian Tissue Transplantation
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.
Introduction In women, approximately one tenth of cancers occur in
