Comparison of the Effect of Cryoprotective Vitrification Agent with Different Concentrations on Protecting 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, Sichuan 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：

Vitrification; Concentration; Human ovarian tissue; Apoptosis

DOI：

10.7507/1002-0179.201600240

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To compare the effect of cryoprotective vitrification agent with different concentrations on protecting human ovarian tissue. Methods Human ovarian biopsy tissues were obtained from nine patients between August 2013 and May 2014. The cortical tissue pieces obtained from each patient were cryopreserved using direct cover vitrification (DCV) with two different concentrations. The vitrification solutions were divided into two groups: high concentration [15% (V/V) dimethyl sulphoxide (DMSO)+15% (V/V) ethylene glycol (EG)+0.5 mol/L sucrose] and low concentration group (12% DMSO+12% EG+0.5 mol/L sucrose). The preservation effects in the two different concentration groups were compared by histologic evaluation using light microscope and apoptosis assessed by terminal dexoynucleotidyl transferase-mediated nick end labeling staining. Results There was no significant difference in the proportion of morphologically normal primordial follicles between high concentration group and low concentration group (P ＞ 0.05) . The proportion of apoptotic primordial follicles in the low concentration group was 29.7% (58/195) , and was 42.1% (69/164) in the high concentration group, with a significant difference between the two groups (P ＜ 0.05) . The proportion of apoptotic stromal cells in the low and high concentration group was 30.2% (162/537) and 41.9% (206/492) respectively with a significant difference (P ＜ 0.05) . Conclusions Vitrification solutions with lower concentration can improve the preservation of the primordial follicles and stromal cells in human ovarian tissue. It is a method worth trying in order to improve the protective effect of vitrification by decreasing the toxicity of vitrification solutions.

Citation： XiaoZhun, ZhangYaoyao, LiLingling, LiShangwei. Comparison of the Effect of Cryoprotective Vitrification Agent with Different Concentrations on Protecting Human Ovarian Tissue. West China Medical Journal, 2016, 31(5): 885-888. doi: 10.7507/1002-0179.201600240 Copy

1.	Amorim CA, Dolmans MM, David A, et al. Vitrification and xenografting of human ovarian tissue[J]. Fertil Steril, 2012, 98 (5) : 1291-1298.
2.	Xiao Z, Li SW, Zhang YY, et al. Niv versus dropping vitrification in cryopreservation of human ovarian tissue[J]. Cryo Letters, 2014, 35 (3) : 226-231.
3.	肖准. 人卵巢组织慢速冷冻保护液中添加抗凋亡剂研究[J]. 华西医学, 2016, 31 (3) : 507-510.
4.	肖准. 新型玻璃化冷冻法对人卵巢组织超微结构保存的研究[J]. 华西医学, 2013, 28 (4) : 567-570.
5.	Xiao Z, Wang Y, Li LL, et al. In vitro culture thawed human ovarian tissue: NIV versus slow freezing method[J]. Cryo Letters, 2013, 34 (2) : 520-526.
6.	Chen SU, Chien CL, Wu MY, et al. Novel direct cover vitrification for cryopreservation of ovarian tissues increases follicle viability and pregnancy capability in mice[J]. Hum Reprod, 2006, 21 (11) : 2794- 2800.
7.	Gougeon A. Dynamics of follicular growth in the human: a model from preliminary results[J]. Hum Reprod, 1986, 1 (2) : 81-87.
8.	Raffaella D, Luigi N, Giuseppe L, et al. Evidence of apoptosis in human primordial and primary follicles[J]. Hum Reprod, 2003, 18 (12) : 2678-2682.
9.	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.
10.	赵硕. 卵巢组织玻璃化冷冻研究进展[J]. 国际生殖健康/计划生育杂志, 2013, 32 (1) : 45-49.
11.	Amorim CA, David A, Van LA, et al. Vtrification of human ovarian tissue: effect of different solutions and procedures[J]. Fertil Steril, 2011, 95 (3) : 1094-1097.
12.	Isachenko E, Isachenko V, Rahimi G, et al. Cryopreservation of human ovarian tissue by direct plunging into liquid Nitrogen[J]. Eur J Obstet Gynecol Reprod Biol, 2003, 108 (2) : 186-193.
13.	Li YB, Zhou CQ, Yang GF, et al. Modified vitrification method for cryopreservation of human ovarian tissues[J]. Chin Med J, 2007, 120 (2) : 110-114.
14.	Huang LL, Mo YQ, Wang WJ, et al. Cryopreservation of human ovarian tissue by solid-surface vitrification[J]. Eur J Obstet Gynecol Reprod Biol, 2008, 139 (2) : 193-198.
15.	Wang Y, Xiao Z, Li L, et al. Novel needle immersed vitrification: a practical and convenient method with potential advantages in mouse and human ovarian tissue cryopreservation[J]. Hum Reprod, 2008, 23 (10) : 2256-2265.
16.	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.

1. Amorim CA, Dolmans MM, David A, et al. Vitrification and xenografting of human ovarian tissue[J]. Fertil Steril, 2012, 98 (5) : 1291-1298.
2. Xiao Z, Li SW, Zhang YY, et al. Niv versus dropping vitrification in cryopreservation of human ovarian tissue[J]. Cryo Letters, 2014, 35 (3) : 226-231.
3. 肖准. 人卵巢组织慢速冷冻保护液中添加抗凋亡剂研究[J]. 华西医学, 2016, 31 (3) : 507-510.
4. 肖准. 新型玻璃化冷冻法对人卵巢组织超微结构保存的研究[J]. 华西医学, 2013, 28 (4) : 567-570.
5. Xiao Z, Wang Y, Li LL, et al. In vitro culture thawed human ovarian tissue: NIV versus slow freezing method[J]. Cryo Letters, 2013, 34 (2) : 520-526.
6. Chen SU, Chien CL, Wu MY, et al. Novel direct cover vitrification for cryopreservation of ovarian tissues increases follicle viability and pregnancy capability in mice[J]. Hum Reprod, 2006, 21 (11) : 2794- 2800.
7. Gougeon A. Dynamics of follicular growth in the human: a model from preliminary results[J]. Hum Reprod, 1986, 1 (2) : 81-87.
8. Raffaella D, Luigi N, Giuseppe L, et al. Evidence of apoptosis in human primordial and primary follicles[J]. Hum Reprod, 2003, 18 (12) : 2678-2682.
9. 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.
10. 赵硕. 卵巢组织玻璃化冷冻研究进展[J]. 国际生殖健康/计划生育杂志, 2013, 32 (1) : 45-49.
11. Amorim CA, David A, Van LA, et al. Vtrification of human ovarian tissue: effect of different solutions and procedures[J]. Fertil Steril, 2011, 95 (3) : 1094-1097.
12. Isachenko E, Isachenko V, Rahimi G, et al. Cryopreservation of human ovarian tissue by direct plunging into liquid Nitrogen[J]. Eur J Obstet Gynecol Reprod Biol, 2003, 108 (2) : 186-193.
13. Li YB, Zhou CQ, Yang GF, et al. Modified vitrification method for cryopreservation of human ovarian tissues[J]. Chin Med J, 2007, 120 (2) : 110-114.
14. Huang LL, Mo YQ, Wang WJ, et al. Cryopreservation of human ovarian tissue by solid-surface vitrification[J]. Eur J Obstet Gynecol Reprod Biol, 2008, 139 (2) : 193-198.
15. Wang Y, Xiao Z, Li L, et al. Novel needle immersed vitrification: a practical and convenient method with potential advantages in mouse and human ovarian tissue cryopreservation[J]. Hum Reprod, 2008, 23 (10) : 2256-2265.
16. 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.

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Comparison of the Effect of Cryoprotective Vitrification Agent with Different Concentrations on Protecting Human Ovarian Tissue

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