Skip to main content

Advertisement

SpringerLink
Log in

VEGF and bFGF increase survival of xenografted human ovarian tissue in an experimental rabbit model

  • Fertility Preservation
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

The aim of this study is to determine whether vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) could increase the survival of xenografted human ovarian tissue in an experimental rabbit model.

Methods

Fresh human ovarian tissue was xenotransplanted into the back muscle of 25 castrated female New Zealand rabbits for 6 weeks with the immunosuppression of FTY720 (2 mg/kg/d). Rabbits were randomly divided into five experimental groups: (A) graft and host treatment with VEGF (50 ng/ml); (B) graft and host treatment with bFGF (100 ng/ml); (C) graft and host treatment with VEGF(50 ng/ml) + bFGF (100 ng/ml); (D) graft and host treatment with normal saline; (E) control group, no treatment. 4 weeks after transplantation, human menopausal gonadotropin (HMG) 10 IU was administered every second day in group A, group B, group C and group D for 2 weeks. Graft survival was assessed by graft recovery rate, histological analysis, immunohistochemical staining for CD31 and Ki-67expression, TUNEL assay.

Results

After 6 weeks of grafting, the number of CD31-positive stained cells increased significantly in group A, group B and group C compared to the control group. All groups showed strong Ki-67 immunostaining in ovarian stroma. Only one rabbit in group C retained the grafts’ follicles. Grafting resulted in relative lower fibrosis in group A and group C compared to the control group. Apoptosis was significantly lower in group C compared to the control group.

Conclusions

Fresh human ovarian cortex grafted into the back muscle of rabbit can sustain part of ovarian tissue function with the immunosuppression of FTY720, although follicle number diminishes significantly after grafting. The administration of VEGF and bFGF, especially the combination of them, may trigger angiogenesis, reduce apoptosis and fibrosis, increase survival in transplanted human ovarian tissue.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. McLaren JF, Bates GW. Fertility preservation in women of reproductive age with cancer. Am J Obstet Gynecol. 2012;207:455–62.

    Article  PubMed  Google Scholar 

  2. Lee SJ, Schover LR, Partridge AH, Patrizio P, Wallace WH, Hagerty K, et al. American Society of Clinical Oncology recommendations on fertility preservation in cancer patients. J Clin Oncol. 2006;24:2917–31.

    Article  PubMed  Google Scholar 

  3. Baka SG, Toth TL, Veeck LL, Jones HJ, Muasher SJ, Lanzendorf SE. Evaluation of the spindle apparatus of in-vitro matured human oocytes following cryopreservation. Hum Reprod. 1995;10:1816–20.

    PubMed  CAS  Google Scholar 

  4. Silber SJ. Ovary cryopreservation and transplantation for fertility preservation. Mol Hum Reprod. 2012;18:59–67.

    Article  PubMed  CAS  Google Scholar 

  5. Donnez J, Jadoul P, Pirard C, Hutchings G, Demylle D, Squifflet J, et al. Live birth after transplantation of frozen-thawed ovarian tissue after bilateral oophorectomy for benign disease. Fertil Steril. 2012;98:720–5.

    Article  PubMed  Google Scholar 

  6. Revelli A, Marchino G, Dolfin E, Molinari E, Delle PL, Salvagno F, et al. Live birth after orthotopic grafting of autologous cryopreserved ovarian tissue and spontaneous conception in Italy. Fertil Steril. 2013;99:227–30.

    Article  PubMed  Google Scholar 

  7. Dolmans MM, Marinescu C, Saussoy P, Van Langendonckt A, Amorim C, Donnez J. Reimplantation of cryopreserved ovarian tissue from patients with acute lymphoblastic leukemia is potentially unsafe. Blood. 2010;116:2908–14.

    Article  PubMed  CAS  Google Scholar 

  8. Nugent D, Newton H, Gallivan L, Gosden RG. Protective effect of vitamin E on ischaemia-reperfusion injury in ovarian grafts. J Reprod Fertil. 1998;114:341–6.

    Article  PubMed  CAS  Google Scholar 

  9. Israely T, Dafni H, Nevo N, Tsafriri A, Neeman M. Angiogenesis in ectopic ovarian xenotransplantation: multiparameter characterization of the neovasculature by dynamic contrast-enhanced MRI. Magn Reson Med. 2004;52:741–50.

    Article  PubMed  Google Scholar 

  10. Van Eyck AS, Jordan BF, Gallez B, Heilier JF, Van Langendonckt A, Donnez J. Electron paramagnetic resonance as a tool to evaluate human ovarian tissue reoxygenation after xenografting. Fertil Steril. 2009;92:374–81.

    Article  PubMed  CAS  Google Scholar 

  11. Demeestere I, Simon P, Emiliani S, Delbaere A, Englert Y. Orthotopic and heterotopic ovarian tissue transplantation. Hum Reprod Update. 2009;15:649–65.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  12. Zhou XH, Wu YJ, Shi J, Xia YX, Zheng SS. Cryopreservation of human ovarian tissue: comparison of novel direct cover vitrification and conventional vitrification. Cryobiology. 2010;60:101–5.

    Article  PubMed  CAS  Google Scholar 

  13. Hazzard TM, Stouffer RL. Angiogenesis in ovarian follicular and luteal development. Baillieres Best Pract Res Clin Obstet Gynaecol. 2000;14:883–900.

    Article  PubMed  CAS  Google Scholar 

  14. Kaczmarek MM, Schams D, Ziecik AJ. Role of vascular endothelial growth factor in ovarian physiology - an overview. Reprod Biol. 2005;5:111–36.

    PubMed  Google Scholar 

  15. Ravindranath N, Little-Ihrig L, Phillips HS, Ferrara N, Zeleznik AJ. Vascular endothelial growth factor messenger ribonucleic acid expression in the primate ovary. Endocrinology. 1992;131:254–60.

    Article  PubMed  CAS  Google Scholar 

  16. Seli E, Zeyneloglu HB, Senturk LM, Bahtiyar OM, Olive DL, Arici A. Basic fibroblast growth factor: peritoneal and follicular fluid levels and its effect on early embryonic development. Fertil Steril. 1998;69:1145–8.

    Article  PubMed  CAS  Google Scholar 

  17. Reynolds LP, Redmer DA. Expression of the angiogenic factors, basic fibroblast growth factor and vascular endothelial growth factor, in the ovary. J Anim Sci. 1998;76:1671–81.

    PubMed  CAS  Google Scholar 

  18. Imthurn B, Cox SL, Jenkin G, Trounson AO, Shaw JM. Gonadotrophin administration can benefit ovarian tissue grafted to the body wall: implications for human ovarian grafting. Mol Cell Endocrinol. 2000;163:141–6.

    Article  PubMed  CAS  Google Scholar 

  19. Chiba K, Yanagawa Y, Masubuchi Y, Kataoka H, Kawaguchi T, Ohtsuki M, et al. FTY720, a novel immunosuppressant, induces sequestration of circulating mature lymphocytes by acceleration of lymphocyte homing in rats. I. FTY720 selectively decreases the number of circulating mature lymphocytes by acceleration of lymphocyte homing. J Immunol. 1998;160:5037–44.

    PubMed  CAS  Google Scholar 

  20. Dath C, Van Eyck AS, Dolmans MM, Romeu L, Delle VL, Donnez J, et al. Xenotransplantation of human ovarian tissue to nude mice: comparison between four grafting sites. Hum Reprod. 2010;25:1734–43.

    Article  PubMed  CAS  Google Scholar 

  21. Meirow D, Hardan I, Dor J, Fridman E, Elizur S, Ra’Anani H, et al. Searching for evidence of disease and malignant cell contamination in ovarian tissue stored from hematologic cancer patients. Hum Reprod. 2008;23:1007–13.

    Article  PubMed  Google Scholar 

  22. Lotz L, Montag M, van der Ven H, von Wolff M, Mueller A, Hoffmann I, et al. Xenotransplantation of cryopreserved ovarian tissue from patients with ovarian tumors into SCID mice–no evidence of malignant cell contamination. Fertil Steril. 2011;95:2612–4.

    Article  PubMed  Google Scholar 

  23. Chambers EL, Gosden RG, Yap C, Picton HM. In situ identification of follicles in ovarian cortex as a tool for quantifying follicle density, viability and developmental potential in strategies to preserve female fertility. Hum Reprod. 2010;25:2559–68.

    Article  PubMed  CAS  Google Scholar 

  24. Amorim CA, Dolmans MM, David A, Jaeger J, Vanacker J, Camboni A, et al. Vitrification and xenografting of human ovarian tissue. Fertil Steril. 2012;98:1291–8.

    Article  PubMed  Google Scholar 

  25. David A, Dolmans MM, Van Langendonckt A, Donnez J, Amorim CA. Immunohistochemical localization of growth factors after cryopreservation and 3 weeks’ xenotransplantation of human ovarian tissue. Fertil Steril. 2011;95:1241–6.

    Article  PubMed  CAS  Google Scholar 

  26. Soleimani R, Heytens E, Oktay K. Enhancement of neoangiogenesis and follicle survival by sphingosine-1-phosphate in human ovarian tissue xenotransplants. PLoS One. 2011;6:e19475.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  27. Friedman O, Orvieto R, Fisch B, Felz C, Freud E, Ben-Haroush A, et al. Possible improvements in human ovarian grafting by various host and graft treatments. Hum Reprod. 2012;27:474–82.

    Article  PubMed  CAS  Google Scholar 

  28. Israely T, Nevo N, Harmelin A, Neeman M, Tsafriri A. Reducing ischaemic damage in rodent ovarian xenografts transplanted into granulation tissue. Hum Reprod. 2006;21:1368–79.

    Article  PubMed  Google Scholar 

  29. Donnez J, Dolmans MM, Demylle D, Jadoul P, Pirard C, Squifflet J, et al. Livebirth after orthotopic transplantation of cryopreserved ovarian tissue. Lancet. 2004;364:1405–10.

    Article  PubMed  CAS  Google Scholar 

  30. Donnez J, Dolmans MM, Demylle D, Jadoul P, Pirard C, Squifflet J, et al. Restoration of ovarian function after orthotopic (intraovarian and periovarian) transplantation of cryopreserved ovarian tissue in a woman treated by bone marrow transplantation for sickle cell anaemia: case report. Hum Reprod. 2006;21:183–8.

    Article  PubMed  CAS  Google Scholar 

  31. Abir R, Fisch B, Jessel S, Felz C, Ben-Haroush A, Orvieto R. Improving posttransplantation survival of human ovarian tissue by treating the host and graft. Fertil Steril. 2011;95:1205–10.

    Article  PubMed  Google Scholar 

  32. Yang H, Lee HH, Lee HC, Ko DS, Kim SS. Assessment of vascular endothelial growth factor expression and apoptosis in the ovarian graft: can exogenous gonadotropin promote angiogenesis after ovarian transplantation? Fertil Steril. 2008;90:1550–8.

    Article  PubMed  CAS  Google Scholar 

  33. Nilsson E, Parrott JA, Skinner MK. Basic fibroblast growth factor induces primordial follicle development and initiates folliculogenesis. Mol Cell Endocrinol. 2001;175:123–30.

    Article  PubMed  CAS  Google Scholar 

  34. Lund EL, Thorsen C, Pedersen MW, Junker N, Kristjansen PE. Relationship between vessel density and expression of vascular endothelial growth factor and basic fibroblast growth factor in small cell lung cancer in vivo and in vitro. Clin Cancer Res. 2000;6:4287–91.

    PubMed  CAS  Google Scholar 

  35. Skinner MK. Regulation of primordial follicle assembly and development. Hum Reprod Update. 2005;11:461–71.

    Article  PubMed  Google Scholar 

  36. Van Eyck AS, Bouzin C, Feron O, Romeu L, Van Langendonckt A, Donnez J, et al. Both host and graft vessels contribute to revascularization of xenografted human ovarian tissue in a murine model. Fertil Steril. 2010;93:1676–85.

    Article  PubMed  Google Scholar 

  37. Tille JC, Wood J, Mandriota SJ, Schnell C, Ferrari S, Mestan J, et al. Vascular endothelial growth factor (VEGF) receptor-2 antagonists inhibit VEGF- and basic fibroblast growth factor-induced angiogenesis in vivo and in vitro. J Pharmacol Exp Ther. 2001;299:1073–85.

    PubMed  CAS  Google Scholar 

  38. Stavri GT, Zachary IC, Baskerville PA, Martin JF, Erusalimsky JD. Basic fibroblast growth factor upregulates the expression of vascular endothelial growth factor in vascular smooth muscle cells. Synergistic interaction with hypoxia. Circulation. 1995;92:11–4.

    Article  PubMed  CAS  Google Scholar 

  39. Mattern J, Koomagi R, Volm M. Coexpression of VEGF and bFGF in human epidermoid lung carcinoma is associated with increased vessel density. Anticancer Res. 1997;17:2249–52.

    PubMed  CAS  Google Scholar 

  40. Shikanov A, Zhang Z, Xu M, Smith RM, Rajan A, Woodruff TK, et al. Fibrin encapsulation and vascular endothelial growth factor delivery promotes ovarian graft survival in mice. Tissue Eng Part A. 2011;17:3095–104.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  41. Laschke MW, Menger MD, Vollmar B. Ovariectomy improves neovascularization and microcirculation of freely transplanted ovarian follicles. J Endocrinol. 2002;172:535–44.

    Article  PubMed  CAS  Google Scholar 

  42. Dissen GA, Lara HE, Fahrenbach WH, Costa ME, Ojeda SR. Immature rat ovaries become revascularized rapidly after autotransplantation and show a gonadotropin-dependent increase in angiogenic factor gene expression. Endocrinology. 1994;134:1146–54.

    Article  PubMed  CAS  Google Scholar 

  43. Soleimani R, Heytens E, Van den Broecke R, Rottiers I, Dhont M, Cuvelier CA, et al. Xenotransplantation of cryopreserved human ovarian tissue into murine back muscle. Hum Reprod. 2010;25:1458–70.

    Article  PubMed  CAS  Google Scholar 

  44. von Wolff M, Donnez J, Hovatta O, Keros V, Maltaris T, Montag M, et al. Cryopreservation and autotransplantation of human ovarian tissue prior to cytotoxic therapy–a technique in its infancy but already successful in fertility preservation. Eur J Cancer. 2009;45:1547–53.

    Article  Google Scholar 

  45. Amorim CA, David A, Dolmans MM, Camboni A, Donnez J, Van Langendonckt A. Impact of freezing and thawing of human ovarian tissue on follicular growth after long-term xenotransplantation. J Assist Reprod Genet. 2011;28:1157–65.

    Article  PubMed  PubMed Central  Google Scholar 

  46. Liu J, Van der Elst J, Van den Broecke R, Dhont M. Live offspring by in vitro fertilization of oocytes from cryopreserved primordial mouse follicles after sequential in vivo transplantation and in vitro maturation. Biol Reprod. 2001;64:171–8.

    Article  PubMed  CAS  Google Scholar 

  47. Fujita T, Hirose R, Yoneta M, Sasaki S, Inoue K, Kiuchi M, et al. Potent immunosuppressants, 2-alkyl-2-aminopropane-1,3-diols. J Med Chem. 1996;39:4451–9.

    Article  PubMed  CAS  Google Scholar 

  48. Yagi H, Kamba R, Chiba K, Soga H, Yaguchi K, Nakamura M, et al. Immunosuppressant FTY720 inhibits thymocyte emigration. Eur J Immunol. 2000;30:1435–44.

    Article  PubMed  CAS  Google Scholar 

  49. Zelinski MB, Murphy MK, Lawson MS, Jurisicova A, Pau KY, Toscano NP, et al. In vivo delivery of FTY720 prevents radiation-induced ovarian failure and infertility in adult female nonhuman primates. Fertil Steril. 2011;95:1440–5.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

Download references

Acknowledgments

This research was supported by the Fund of National Science and Technology Support Plan (NO.2012BAI32B04).

Author information

Authors and Affiliations

  1. Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou, 310006, China

    Lin Wang, Ying-fen Ying, Yin-luan Ouyang, Jing-fen Wang & Jian Xu

Authors
  1. Lin Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ying-fen Ying
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yin-luan Ouyang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jing-fen Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jian Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jian Xu.

Additional information

Capsule The administration of VEGF and bFGF may trigger angiogenesis, reduce apoptosis and fibrosis, increase survival in human ovarian tissue xenotransplantation.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, L., Ying, Yf., Ouyang, Yl. et al. VEGF and bFGF increase survival of xenografted human ovarian tissue in an experimental rabbit model. J Assist Reprod Genet 30, 1301–1311 (2013). https://doi.org/10.1007/s10815-013-0043-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-013-0043-9

Keywords

Search

Navigation