Du Tianqi, Chao Lan, Zhao Shuqin, Chi Linglong, Li Dong, Shen Yanjun, Shi Qing, Deng Xiaohui
Center for Reproduction and Genetics, Suzhou Hospital Affiliated to Nanjing Medical University, 26 Daoqian Street, Suzhou, Jiangsu, 215002, China,
J Assist Reprod Genet. 2015 Aug;32(8):1267-75. doi: 10.1007/s10815-015-0513-3. Epub 2015 Jun 19.
The study aims to assess the protective effects of dimethyl sulfoxide (DMSO)-free solution based on trehalose on the cryopreservation of a whole sheep ovary and evaluate its use as an efficient cryoprotectant.
Twenty-one ovaries collected from 6- to 8-month-old non-pregnant female sheep were randomly distributed into three groups, namely, a fresh group, a DMSO-free group, and a DMSO group. The morphology, cell apoptosis (by hematoxylin and eosin (HE) staining and terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) assay), and mRNA transcript of Bcl-2-associated X protein (BAX) and cold inducible RNA-binding protein (CIRP) (by real-time PCR) of the thawed sheep ovaries and fresh controls were tested to establish a criterion for appraising the results of the cryopreservation.
(i) The histological assessment indicated that the structure of the DMSO-free ovaries remained largely intact and comparable to those of the fresh control groups; whereas, significant damage was observed in the ovaries of the DMSO group (P < 0.05). (ii) The TUNEL assay and mRNA transcript of the BAX assessment showed that the apoptosis parameter in the fresh group was the lowest among all the groups (P < 0.05), and the parameter in the DMSO-free group was significantly lower than that in the DMSO group (P < 0.05). (iii) The level of the CIRP transcripts increased the most in the DMSO-free group followed by the DMSO group and the fresh control group (P < 0.05).
These results indicate that a DMSO-free cryoprotectant solution, especially a trehalose cryoprotectant, is an efficient cryoprotectant and has a beneficial effect on the cryopreservation of whole sheep ovaries.
本研究旨在评估基于海藻糖的无二甲基亚砜（DMSO）溶液对整个绵羊卵巢冷冻保存的保护作用，并评估其作为一种高效冷冻保护剂的用途。
从6至8月龄未怀孕的雌性绵羊收集的21个卵巢随机分为三组，即新鲜组、无DMSO组和DMSO组。对解冻后的绵羊卵巢和新鲜对照组的形态、细胞凋亡（通过苏木精和伊红（HE）染色以及末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）测定）以及Bcl-2相关X蛋白（BAX）和冷诱导RNA结合蛋白（CIRP）的mRNA转录本（通过实时PCR）进行检测，以建立评估冷冻保存结果的标准。
（i）组织学评估表明，无DMSO组卵巢的结构基本保持完整，与新鲜对照组相当；而DMSO组卵巢则观察到明显损伤（P<0.05）。（ii）TUNEL测定和BAX评估的mRNA转录本显示，新鲜组的凋亡参数在所有组中最低（P<0.05），无DMSO组的参数显著低于DMSO组（P<0.05）。（iii）CIRP转录本水平在无DMSO组中增加最多，其次是DMSO组和新鲜对照组（P<0.05）。
这些结果表明，无DMSO冷冻保护剂溶液，尤其是海藻糖冷冻保护剂，是一种高效的冷冻保护剂，对整个绵羊卵巢的冷冻保存具有有益作用。
