The improved protective efficacy of combined antioxidants on cryopreserved and thawed human ovarian tissue

doi:10.3969/j.issn.1006-7795.2021.04.004

Abstract

Abstract: Objective To investigate the protective efficacy of antioxidants N-acetyl-L-cysteine(NAC) and taurine(T) based on the currently used human ovarian tissue cryopreservation and thawing protocol in Beijing Obstetrics and Gynecology Hospital, Capital Medical University, in order to better preserve the viability of follicles and stroma cells, and improve the folliculogenesis potential. Methods The currently used cryopreservation and thawing protocol in our center was set as control group (group C), with addition of 5 mmol NAC (group N) or 0.5 mmol taurine (group T), or with addition of 5 mmol NAC and 0.5 mmol taurine (group N+T). Ovarian tissues from 16 patients were assigned into the 4 groups randomly. The biopsies were cryopreserved and thawed according to the standard operation procedure in our center. After thawing, HE staining and Calcein-AM staining were conducted to evaluate the morphological characteristics and follicle viability. After cultured in vitro for 4 days, the 17β-estradiol (E2) and anti-mullerian hormone (AMH)were measured, and the level of cellular reactive oxygen species (ROS) was also detected. Results All 4 groups obtained good preservation of follicle integrity and viability without statistical significance(P>0.05). After 4 days culture, E2 concentration in culture medium showed no significant difference in 4 groups (P>0.05), AMH concentration in group N+T were significantly higher than that in group C (P<0.05). Group N and Group N+T had lower ROS levels than group C (P<0.05). Conclusions The addition of NAC alone or taurine alone or combination of NAC and taurine could preserve the integrity and viability of the follicle. NAC and taurine could act synergistically to increase the level of E2 and AMH and improve the overall activity of ovarian tissues after thawing, decrease intracellular ROS production,lower the level of oxidative stress, and improve the potential development of follicles after ovarian tissue thawing.

Key words: antioxidant, follicle viability, follicle morphology, follicle development, oxidative stress

CLC Number:

Li Yanglu, Ruan Xiangyan, Cheng Jiaojiao, Zhou Qi, Du Juan, Jin Fengyu, Gu Muqing. The improved protective efficacy of combined antioxidants on cryopreserved and thawed human ovarian tissue[J]. Journal of Capital Medical University, 2021, 42(4): 526-532.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2021.04.004

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2021/V42/I4/526

References

[1] Dolmans M M, Falcone T, Patrizio P. Importance of patient selection to analyze in vitro fertilization outcome with transplanted cryopreserved ovarian tissue[J]. Fertil Steril, 2020, 114(2): 279-280.
[2] 阮祥燕,杜娟,卢丹,等. 中国自体卵巢组织冻存移植后成功妊娠首例报道[J]. 中国临床医生杂志,2021,49(3): 375-378.
[3] Practice Commitlee of the American Society for Reproductive Medicine. Fertility preservation in patients undergoing gonadotoxic therapy or gonadectomy: a committee opinion[J]. Fertil Steril, 2019, 112(6): 1022-1033.
[4] Lee J, Kong H S, Kim E J, et al. Ovarian injury during cryopreservation and transplantation in mice: a comparative study between cryoinjury and ischemic injury[J]. Hum Reprod, 2016, 31(8): 1827-1837.
[5] Gorricho C M, Tavares M R, Apparicio M, et al. Vitrification of cat ovarian tissue: does fragment size matters?[J]. Reprod Domest Anim, 2018, 53 Suppl 3: 125-132.
[6] Max M C, Bizarro-Silva C, Bufalo I, et al. In vitro culture supplementation of EGF for improving the survival of equine preantral follicles[J]. In Vitro Cell Dev Biol Anim, 2018, 54(10): 687-691.
[7] Song Y, Wang H, Huang H, et al. Comparison of the efficacy between NAC and metformin in treating PCOS patients: a meta-analysis[J]. Gynecol Endocrinol, 2020, 36(3): 204-210.
[8] Mu T, Yang J, Li Z, et al. Effect of taurine on reproductive hormone secretion in female rats[J]. Adv Exp Med Biol, 2015, 803: 449-456.
[9] Sanfilippo S, Canis M, Romero S, et al. Quality and functionality of human ovarian tissue after cryopreservation using an original slow freezing procedure[J]. J Assist Reprod Genet, 2013, 30(1): 25-34.
[10] Li Y, Ruan X, Liebenthron J, et al. Ovarian tissue cryopreservation for patients with premature ovary insufficiency caused by cancer treatment: optimal protocol[J]. Climacteric, 2019, 22(4):1-7.
[11] 国际妇科内分泌学会中国妇科内分泌学分会及共识专家. 卵巢组织冻存与移植中国专家共识[J]. 中国临床医生杂志,2018,46(4): 496-500.
[12] Gougeon A. Regulation of ovarian follicular development in primates: facts and hypotheses[J]. Endocr Rev, 1996, 17(2): 121-155.
[13] McLaughlin M, Kinnell H L, Anderson R A, et al. Inhibition of phosphatase and tensin homologue (PTEN) in human ovary in vitro results in increased activation of primordial follicles but compromises development of growing follicles[J]. Mol Hum Reprod, 2014, 20(8): 736-744.
[14] Gougeon A. Dynamics of follicular growth in the human: a model from preliminary results[J]. Hum Reprod, 1986, 1(2): 81-87.
[15] Youm H W, Lee J R, Lee J, et al. Optimal vitrification protocol for mouse ovarian tissue cryopreservation: effect of cryoprotective agents and in vitro culture on vitrified-warmed ovarian tissue survival[J]. Hum Reprod, 2014, 29(4): 720-730.
[16] Herraiz S, Novella-Maestre E, Rodriguez B, et al. Improving ovarian tissue cryopreservation for oncologic patients: slow freezing versus vitrification, effect of different procedures and devices[J]. Fertil Steril, 2014, 101(3): 775-784.
[17] Bastings L, Liebenthron J, Westphal J R, et al. Efficacy of ovarian tissue cryopreservation in a major European center[J]. J Assist Reprod Genet, 2014, 31(8): 1003-1012.
[18] Mahmoodi M, Soleimani M M, Shariatzadeh S M, et al. N-acetylcysteine improves function and follicular survival in mice ovarian grafts through inhibition of oxidative stress[J]. Reprod Biomed Online, 2015, 30(1): 101-110.
[19] Schmidt K L, Byskov A G, Nyboe Andersen A, et al. Density and distribution of primordial follicles in single pieces of cortex from 21 patients and in individual pieces of cortex from three entire human ovaries[J]. Hum Reprod, 2003, 18(6): 1158-1164.
[20] Ramezani M, Salehnia M, Jafarabadi M. Short term culture of vitrified human ovarian cortical tissue to assess the cryopreservation outcome: molecular and morphological analysis[J]. J Reprod Infertil, 2017, 18(1): 162-171.
[21] Kano M, Sosulski A E, Zhang L, et al. AMH/MIS as a contraceptive that protects the ovarian reserve during chemotherapy[J]. Proc Natl Acad Sci U S A, 2017, 114(9): E1688-E1697.
[22] Pepin D, Sabatini M E, Donahoe P K. Mullerian inhibiting substance/anti-Mullerian hormone as a fertility preservation agent[J]. Curr Opin Endocrinol Diabetes Obes, 2018, 25(6): 399-405.
[23] Gerritse R, Beerendonk C C, Westphal J R, et al. Glucose/lactate metabolism of cryopreserved intact bovine ovaries as a novel quantitative marker to assess tissue cryodamage[J]. Reprod Biomed Online, 2011, 23(6): 755-764.
[24] Isachenko V, Todorov P, Isachenko E, et al. Long-Time cooling before cryopreservation decreased translocation of phosphatidylserine (Ptd-L-Ser) in human ovarian tissue[J]. PLoS One, 2015, 10(6): e129108.
[25] Gao J M, Yan J, Li R, et al. Improvement in the quality of heterotopic allotransplanted mouse ovarian tissues with basic fibroblast growth factor and fibrin hydrogel[J]. Hum Reprod, 2013, 28(10): 2784-2793.