猪卵巢组织玻璃化冷冻保存初步研究

为了建立一种有效的卵巢组织玻璃化冷冻方法,试验采用乙二醇(EG) 二甲基亚矾(DMSO)作为玻璃化冷冻保护剂,将猪卵巢组织经过不同浓度的平衡液中处理后进行玻璃化冷冻保存。结果表明,经过两步平衡冷冻组卵巢组织中的卵泡形态结构基本正常;提取卵巢组织细胞DNA进行琼脂糖凝胶电泳,检测玻璃化冷冻卵巢组织细胞DNA损伤,各实验组与对照组的结果无显著差异。显示采用EG DMSO作为玻璃化冷冻保护剂、两步平衡玻璃化冷冻方法可用于猪卵巢组织的冷冻保存,该方法不造成卵巢组织细胞DNA的损伤。

Primary Study on Vitrification of Porcine Ovarian Tissues

In order to establish an efficient vitrification of ovarian tissues. Porcine ovarian tissues were vitrified by being treated with different pre-equilibrated treats, and then being exposed in the vitrification solution with the same time, then being plunged into the liquid nitrogen directly, and using ethylene glycol (EG) and dimethylsulfoxide(DMSO) as cryoprectant. Results shows after vitrification and thawing, the follicle in the surface layer of the ovarian cortex which being treated with two steps of pre-equilibrated remains normal morphology. The DNA in the agarose gel of all research groups were similar as the fresh-control group. Conclusion: The vitrification which uses ethylene glycol (EG) and dimethylsulfoxide(DMSO)as cryoprectant , with two steps of pre-equilibrated and then has been directly plunged into liquid nitrogen may be an efficient cryopreservation method of ovarian cortex fragment. This method didn't injury DNA of the cells.