兔卵母细胞体外成熟、体外受精及受精卵体外培养条件的研究

IVM,IVF of Follicular Oocytes and IVC of in Vitro Fertilized Oocytes in Rabbit

为了建立一个完善的兔胚胎体外生产体系,探讨了TCM-199、M16和KSOM基础培养液对兔卵母细胞体外成熟,上游法和直接洗涤法洗涤处理精液的方法对卵母细胞体外受精及不同浓度的L-谷氨酰胺对受精卵发育的影响.结果表明：以TCM-199为基础培养液体外成熟率（72.29％）显著高于M16成熟率（59.11％）和KSOM成熟率（49.57％）,三者之间彼此差异显著（P＜0.05）;上游法处理精子后,体外受精卵母细胞的卵裂率（64.89％）和囊胚发育率（18.56％）均高于直接洗涤法（51.9％,11.97％）,且差异显著（P ＜0.05）;0.5mmol·L-1L-谷氨酰胺受精卵培养液组的2～4细胞发育率（46.55％）、4～8细胞发育率（39.20％） 、8～16细胞发育率（30.40％）、桑葚胚发育率（26.65％）,均显著高于含1mmol·L-1 L-谷氨酰胺受精卵培养液组（分别为38.83％ 、32.02％ 、25.14％和22.09％）和含2mmol·L-1 L-谷氨酰胺受精卵培养液组（分为33.96％、26.70％、21.43％和17.67％）,且差异显著（P＜0.05）,而含0.5mmol· L-1 L-谷氨酰胺受精卵培养液组囊胚发育率（15.52％）与含1mmol·L-1、2 mmol·L-1 L-谷氨酰胺受精卵培养液组囊胚发育率（分别为10.59％和7.17％）差异显著（P＜0.05）.

In order to establish a perfect in vitro production system of rabbit embryo, We explored the effects of the TCM-199, M16 and KSOM three different basic nutrient solutions on the in vitro maturation of rabbit oocytes, and the upstream method and direct washing method of sperm washing processing methods treating semen of rabbit oocyte in vitro fertilization and different concentrations of L-glutamine on the influence of the rabbit zygote. The results showed that TCM-199 as the foundation nutrient solution in vitro maturation rate （72.29%） was statistically higher than that in the M16 mature rate （59.11%） and KSOM mature rate （49.57%）（p 〈 0.05）. Upstream method processing sperm in vitro fertilized oocytes mother cell of the cleavage rate （64.89%） and segmentation growth rate （18.56%） were higher than direct washing method （51.9%, 11.97%）（/9〈0.05）. 0.5mmol·L^-1 L- glutamine fertilized oocytes nutrient solution 2-4 cell growth rate （46.55%）,4-8 cell growth rate （39.20%）,8-16 cell growth rate （30.40%）,mulberry embryo growth rate （26.65%）,were significantly higher than those with 1 mmol·L^-1 L-glutamine fertilized oocytes nutrient solution （were 38.83%,32.02%,25.14%,22.09%） and 2 mmol·L^-1 L-glutamine fertilized oocytes nutrient solution group （divided into 33.96%,26.70%,21.43%,17.67%）（p 〈 0.05）,and the content of 0.5 mmol·L^-1 L-glutamine fertilized oocytes nutrient solution set of blastula growth rate （15.52%） and with 1 mmol·L^-1,2 mmol·L^-1 L-glutamine fertilized oocytes nutrient solution blastula growth rate respectively （10.59%, 7.17%）（P〈 0.05）.