Influence of different cryopreservation schemes on human ovarian tissue activity

doi:10.3969/j.issn.2095-4344.2013.53.005

Abstract

Abstract:

BACKGROUND:Ovarian tissue cryopreservation is considered to be a safe and effective method to save female reproductive endocrine function, but there is no uniform scheme identified.

OBJECTIVE:To explore the effects of three freezing schemes on human ovarian tissue and the influence on follicle activity.

METHODS: Twenty specimens of human ovarian tissue were frozen by propanediol slow freezing method, dimethyl sulfoxide vitrification method, and direct liquid nitrogen method, respectively. After cell resuscitation, the number of activated cells was counted using the live/dead fluorescence analysis, and estradiol level and follicles at different development stages were measured and counted respectively after cultured in vitro to evaluate the influence of three freezing schemes on human ovarian tissue activity .

RESULTS AND CONCLUSION: The activated oocyte rates of three frozen-thawed ovarian tissue groups were lower than that of the fresh ovarian tissue group (P < 0.05). Among the three frozen-thawed groups, the activated oocyte rate of the dimethyl sulfoxide group was the lowest, and there was no significant difference between the other two groups. After in vitro culture, the estradiol level of the dimethyl sulfoxide group was lower than that in the propanediol and liquid nitrogen groups at 4 days (P < 0.05), and the estradiol levels showed no differences between the fresh ovarian tissue group and three frozen-thawed groups at 8 days. After 14 days of culture, the ratio of growing follicles was increased, but primordial follicles were still in the lead. The total number of normal occytes in the fresh ovarian tissue was higher than that in the three frozen-thawed groups (P < 0.05), and the number of normal occytes in dimethyl sulfoxide group was lower than that in the other two frozen-thawed groups (P < 0.05). These findings indicate that the cryopreservation can cause damage to ovarian tissue follicles but still can save most primordium follicle activity. The primordium follicle cultured in vitro can further develop to have secretion function. The dimethyl sulfoxide vitrification method is superior to the propanediol slow freezing method and direct liquid nitrogen method, but the direct liquid nitrogen method is simple and convenient with stable cryopreservation effects.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: ovary, cryopreservation, cell survival, cell death, ovarian follicle

CLC Number:

R318

Li Yun-xiu, Sun Ying-pu, Tang Li, Ma Yan-ping. Influence of different cryopreservation schemes on human ovarian tissue activity[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(53): 9125-9131.

Figures/Tables 3

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