Investigation on the Effect of Adding Antioxidant Supplementation into the Cryoprotectant Solution on Slow Freezing of Human Ovarian Tissue_West China Medical Journal

Authors：

 XiaoZhun ¹ , ZhangYaoyao ¹ , LiLingling ² ,  LiShangwei ¹

1. Reproductive Medical Center, West China Second University Hospital, Sichuan University, Chengdu 610041, P. R. China;
2. Department of Obstetrics and Gynecology, Sichuan Provincial Hospital for Women and Children, Chengdu, Sichuan 610031, P. R. China;

Corresponding author：

LiShangwei, Email: lishangwei2015@sina.com

Keywords：

Slow freezing; Human ovarian tissue; Apoptosis; Cryoprotectant

DOI：

10.7507/1002-0179.201600134

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To determine if the cryoprotectant solution supplementation with Vitamin C can improve the protective effect of human ovarian tissue cryopreservation by anti-apoptosis. Methods Human ovarian cortical tissues were collected from nine patients treated between March 2012 and April 2013. The cortical tissue pieces obtained from each patient were divided into two groups:control (conventional slow freezing) and trial group (slow freezing supplementation with Vitamin C). The preservation effects in the two groups were compared by histology using light microscope and apoptosis assessed by TUNEL assay. Results The proportion of morphologically normal primordial follicles in the trial group was higher than that in the control group (P<0.05). The proportion of apoptotic primordial follicles and stromal cells in the trial group was lower than that in the control group (P<0.05). Conclusion Cryoprotectant solutions supplementation with Vitamin C can improve the preservation of primordial follicles and stromal cells. It might be a method worth to try in order to improve the protective effect of human ovarian tissue cryopreservation by inhibiting of apoptosis.

Citation： XiaoZhun, ZhangYaoyao, LiLingling, LiShangwei. Investigation on the Effect of Adding Antioxidant Supplementation into the Cryoprotectant Solution on Slow Freezing of Human Ovarian Tissue. West China Medical Journal, 2016, 31(3): 507-510. doi: 10.7507/1002-0179.201600134 Copy

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2.	Poirot C, Abirached F, Prades M, et al. Induction of puberty by autograft of cryopreserved ovarian tissue[J]. Lancet, 2012, 379(9815):588.
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7.	赵婷, 叶大风. 女性癌症患者卵巢组织冻存库的研究现状[J]. 中国妇幼健康研究, 2006, 17(6):544-546.
8.	Gosden RG, Baird DT, Wade JC, et al. Restoration of fertility to oophorectomized sheep by ovarian autografts stored at -196 degrees C[J]. Hum Reprod, 1994, 9(4):597-603.
9.	Gougeon A. Dynamics of follicular growth in the human:a model from preliminary results[J]. Hum Reprod, 1986, 1(2):81-87.
10.	Depalo R, Nappi L, Loverro G, et al. Evidence of apoptosis in human primordial and primary follicles[J]. Hum Reprod, 2003, 18(12):2678-2682.
11.	Kim SS, Yang HW, Kang HG, et al. Quantitative assessment of ischemic tissue damage in ovarian cortical tissue with or without antioxidant (ascorbic acid) treatment[J]. Fertil Steril, 2004, 82(3):679-685.
12.	Chang HJ, Moon JH, Lee JR, et al. Optimal condition of vitrification method for cryopreservation of human ovarian cortical tissues[J]. J Obstet Gynaecol Res, 2011, 37(8):1092-1101.
13.	Xiao Z, Wang Y, Li L, et al. Needle immersed vitrification can lower the concentration of cryoprotectant in human ovarian tissue cryopreservation[J]. Fertil Steril, 2010, 94(6):2323-2328.
14.	Keros V, Xella S, Hultenby K, et al. Vitrification versus controlled-rate freezing in cryopreservation of human ovarian tissue[J]. Human Reproduction, 2009, 24(7):1670-1683.
15.	Rimon E, Cohen T, Dantes A, et al. Apoptosis in cryopreserved human ovarian tissue obtained from cancer patients:a tool for evaluating cryopreservation utility[J]. Int J Oncol, 2005, 27(2):345-353.
16.	Perez GI, Robles R, Knudson CM, et al. Prolongation of ovarian lifespan into advanced chronological age by Bax-deficiency[J]. Nat Genet, 1999, 21(2):200-203.
17.	Xiao Z, Wang Y, Li L, et al. Cryopreservation of the human ovarian tissue induces the expression of Fas system in morphologically normal primordial follicles[J]. Cryo Letters, 2010, 31(2):112-119.
18.	Abir R, Fisch B, Jessel S, et al. Improving posttransplantation survival of human ovarian tissue by treating the host and graft[J]. Fertil Steril, 2011, 95(4):1205-1210.

1. Andersen CY, Kristensen SG, Greve T, et al. Cryopreservation of ovarian tissue for fertility preservation in young female oncological patients[J]. Future Oncol, 2012, 8(5):595-608.
2. Poirot C, Abirached F, Prades M, et al. Induction of puberty by autograft of cryopreserved ovarian tissue[J]. Lancet, 2012, 379(9815):588.
3. Donnez J, Dolmans MM, Demylle D, et al. Livebirth after orthotopic transplantation of cryopreserved ovarian tissue[J]. Lancet, 2004, 364(9443):1405-1410.
4. Ernst E, Bergholdt S, Jørgensen JS, et al. The first woman to give birth to two children following transplantation of frozen/thawed ovarian tissue[J]. Hum Reprod, 2010, 25(5):1280-1281.
5. Donnez J, Squifflet J, Jadoul P, et al. Pregnancy and live birth after autotransplantation of frozen-thawed ovarian tissue in a patient with metastatic disease undergoing chemotherapy and hematopoietic stem cell transplantation[J]. Fertil Steril, 2011, 95(5):1787.e1-1787.e4.
6. Isachenko V, Orth I, Isachenko E, et al. Viability of human ovarian tissue confirmed 5 years after freezing with spontaneous ice-formation by autografting and chorio-allantoic membrane culture[J]. Cryobiology, 2013, 66(3):233-238.
7. 赵婷, 叶大风. 女性癌症患者卵巢组织冻存库的研究现状[J]. 中国妇幼健康研究, 2006, 17(6):544-546.
8. Gosden RG, Baird DT, Wade JC, et al. Restoration of fertility to oophorectomized sheep by ovarian autografts stored at -196 degrees C[J]. Hum Reprod, 1994, 9(4):597-603.
9. Gougeon A. Dynamics of follicular growth in the human:a model from preliminary results[J]. Hum Reprod, 1986, 1(2):81-87.
10. Depalo R, Nappi L, Loverro G, et al. Evidence of apoptosis in human primordial and primary follicles[J]. Hum Reprod, 2003, 18(12):2678-2682.
11. Kim SS, Yang HW, Kang HG, et al. Quantitative assessment of ischemic tissue damage in ovarian cortical tissue with or without antioxidant (ascorbic acid) treatment[J]. Fertil Steril, 2004, 82(3):679-685.
12. Chang HJ, Moon JH, Lee JR, et al. Optimal condition of vitrification method for cryopreservation of human ovarian cortical tissues[J]. J Obstet Gynaecol Res, 2011, 37(8):1092-1101.
13. Xiao Z, Wang Y, Li L, et al. Needle immersed vitrification can lower the concentration of cryoprotectant in human ovarian tissue cryopreservation[J]. Fertil Steril, 2010, 94(6):2323-2328.
14. Keros V, Xella S, Hultenby K, et al. Vitrification versus controlled-rate freezing in cryopreservation of human ovarian tissue[J]. Human Reproduction, 2009, 24(7):1670-1683.
15. Rimon E, Cohen T, Dantes A, et al. Apoptosis in cryopreserved human ovarian tissue obtained from cancer patients:a tool for evaluating cryopreservation utility[J]. Int J Oncol, 2005, 27(2):345-353.
16. Perez GI, Robles R, Knudson CM, et al. Prolongation of ovarian lifespan into advanced chronological age by Bax-deficiency[J]. Nat Genet, 1999, 21(2):200-203.
17. Xiao Z, Wang Y, Li L, et al. Cryopreservation of the human ovarian tissue induces the expression of Fas system in morphologically normal primordial follicles[J]. Cryo Letters, 2010, 31(2):112-119.
18. Abir R, Fisch B, Jessel S, et al. Improving posttransplantation survival of human ovarian tissue by treating the host and graft[J]. Fertil Steril, 2011, 95(4):1205-1210.

West China Medical Journal

Investigation on the Effect of Adding Antioxidant Supplementation into the Cryoprotectant Solution on Slow Freezing of Human Ovarian Tissue

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