2种化学激活方法制备的小鼠孤雌胚胎早期发育指标比较研究

Comparison of the Early Development of Murine Parthenogenetic Embryos Prepared by 2 Different Chemical Activation Methods

为了寻找一种健康、有效的制备孤雌胚胎的方法,从根本上提高哺乳动物孤雌胚胎、体外受精胚胎及核移植胚胎成活率并减少孤雌胚胎、核移植胚胎出生后发生潜在疾病与缺陷的几率。通过比较2种不同化学激活方法(试验组I:7%乙醇+2mmol/L 6DMAP;试验组II:7%乙醇+2mmol/L 6DMAP+5μg/mL CB)制备的小鼠孤雌胚胎由卵母细胞激活到发育至囊胚的时间、囊胚形态及囊胚细胞数目,初步研究了不同化学方法制备的小鼠孤雌胚胎在形态学及细胞水平上的差异性。结果表明:试验组I与试验组II相比,前者比后者卵裂率稍高,桑葚率、不同时间段的囊胚率稍低,但两组间数据差异不显著(64.9vs73.7,P>0.1;31.7vs28.8,P>0.1;8.7vs5.5,P>0.1)。2种方法所得的孤雌囊胚在形态上相似,但是试验组II囊胚细胞数要比试验组I囊胚细胞数稍少(67vs50,P>0.1);两组囊胚的细胞数与受精囊胚相比没有显著差异(67vs50vs65,P>0.1)。选用的2种化学方法制备的小鼠孤雌胚胎,在由卵母细胞发育至囊胚的时间、囊胚形态及囊胚细胞数目方面没有显著差异。

This study was to investigate the better suitable method for making more healthy mouse parthenogenetic embryos for generating mammalian parthenogenetic embryos, in vitro fertilized embryos and nuclear transferred embryos. In this experiment, we using 2 different chemical activation methods to product mouse parthenogenetic embryos. And we analyzed the time from oocytes actviation to blastocystes, the shape and the total cell number of blastoeystes of two group. Group I was treated by 7% alcohol＋2 mmol/L 6DMAP, and group II was by 7% alcohol＋2 mmol/L 6DMAP＋5 μg/mL CB. The results showed that： the cleavage rate of parthenogenetic embryos of Gruop I was higher than that of Group Ⅱ, and the morulae rate and blastocyst rate during different durations was lower than those of Gruop Ⅱ, but there were no significant differences between them （64.9 vs 73.7, P〉0.1; 31.7 vs 28.8, P〉0.1; 8.7 vs 5.5, P〉0.1）. The shape of blastocystes of those 2 groups was similar, but the cell number of blastocystes of Gruop Ⅱ was more than those in Group I （67 vs 50, P〉0.1）,as well as the cell number of blastocyst between these two groups （67 vs 50 vs 65,P〉0.1）. There were no significant differences between 2 different chemical activation groups in the time from oocytes actviation to blastocystes, the shape and the total cell number of blastocystes.