人卵母细胞慢速程序冷冻法冻融技术的初步研究
发布时间：2019-04-15 19:26
【摘要】：目的 本研究设计慢速程序冷冻法冻融人成熟卵母细胞(MII期)与未成熟卵母细胞(GV/MI期),分析冻融后卵母细胞的发育潜能及影响卵母细胞冻融后发育潜能的原因,进而确立“冷冻程序—卵母细胞所处阶段—受精能力及早期胚胎的发育能力”三者间的联系,旨在探索出一条适合人卵母细胞的冻融技术。 方法 将来源于行IVF-ET患者捐赠的67枚多余成熟卵母细胞(MII期)随机分为A、B、C三组,在不同条件下进行慢速冷冻法冻融。根据CPA及解冻液中不同的SSS浓度和不同的去除卵母细胞周围部分颗粒细胞的方法将卵母细胞分组如下:A组:24枚,15%SSS、透明质酸酶消化法去颗粒;B组:21枚,30%SSS、透明质酸酶消化法去颗粒;C组:22枚,30%SSS、机械法去颗粒。另随机获取同时期行IVF-ET中的新鲜卵母细胞(未冷冻)25枚作为对照。比较SSS浓度和去颗粒方法的不同对冻融后的卵母细胞存活率、受精能力及胚胎早期发育力的影响。 将来源于手术中卵巢组织的82枚人未成熟卵母细胞(GV/MI期)随机分为两组,冻融组57枚,新鲜对照组25枚。将冻融组卵母细胞慢速冷冻,冻融后存
[Abstract]:Objective to design a slow programmed freezing and thawing method for human mature oocytes (MII phase) and immature oocytes (GV/MI phase). This paper analyzes the developmental potential of oocytes after freezing and thawing and the reasons that affect the developmental potential of oocytes after freezing and thawing, and then establishes the relationship among "freezing procedure-stage of oocytes-fertilization ability and developmental ability of early embryos". In order to explore a suitable freezing and thawing technology for human oocytes. Methods 67 superfluous mature oocytes (MII phase) from IVF-ET patients were randomly divided into three groups: group A, group B, and group C, which were frozen and thawed by slow freezing under different conditions. According to the different concentration of SSS in CPA solution and different methods of removing granulosa cells around oocytes, oocytes were divided into the following groups: group A: 24 cells, 15% SSS, degranulation by hyaluronidase digestion; Group B: 21, 30% SSS, hyaluronidase digestion degranulation, group C: 22, 30% SSS, mechanical degranulation. In addition, 25 fresh oocytes (unfrozen) from simultaneous IVF-ET were randomly obtained as control. The effects of different concentrations of SSS and degranulation methods on the survival rate, fertilization ability and early embryo development ability of frozen-thawed oocytes were compared. In the future, 82 human immature oocytes (GV/MI phase) derived from ovarian tissue during operation were randomly divided into two groups: freeze-thaw group (57) and fresh control group (25). The oocytes in the freeze-thaw group were frozen slowly and stored after freezing and thawing.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R321
本文编号：2458422
[Abstract]:Objective to design a slow programmed freezing and thawing method for human mature oocytes (MII phase) and immature oocytes (GV/MI phase). This paper analyzes the developmental potential of oocytes after freezing and thawing and the reasons that affect the developmental potential of oocytes after freezing and thawing, and then establishes the relationship among "freezing procedure-stage of oocytes-fertilization ability and developmental ability of early embryos". In order to explore a suitable freezing and thawing technology for human oocytes. Methods 67 superfluous mature oocytes (MII phase) from IVF-ET patients were randomly divided into three groups: group A, group B, and group C, which were frozen and thawed by slow freezing under different conditions. According to the different concentration of SSS in CPA solution and different methods of removing granulosa cells around oocytes, oocytes were divided into the following groups: group A: 24 cells, 15% SSS, degranulation by hyaluronidase digestion; Group B: 21, 30% SSS, hyaluronidase digestion degranulation, group C: 22, 30% SSS, mechanical degranulation. In addition, 25 fresh oocytes (unfrozen) from simultaneous IVF-ET were randomly obtained as control. The effects of different concentrations of SSS and degranulation methods on the survival rate, fertilization ability and early embryo development ability of frozen-thawed oocytes were compared. In the future, 82 human immature oocytes (GV/MI phase) derived from ovarian tissue during operation were randomly divided into two groups: freeze-thaw group (57) and fresh control group (25). The oocytes in the freeze-thaw group were frozen slowly and stored after freezing and thawing.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R321
【引证文献】
相关硕士学位论文 前1条
1 邢琼;雌性配子与人类三原核囊胚的玻璃化冷冻研究[D];安徽医科大学;2007年
，本文编号：2458422
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