植入后全胚胎培养的培养基pH值改变对大鼠胚胎生长发育的影响

目的　探讨植入后全胚胎培养培养基最适pH值。方法　分离 9.5天龄Wistar大鼠胚胎 ,分别在实验组 (即不同pH :5 .5 ,6 .0 ,6 .5 ,7.0 ,7.5 ,8.0 ,8.5 ,9.0 )和对照组 (pH为 8.2 )的培养基 (1 0 0 %大鼠即刻离心血清 )中体外旋转培养4 8小时 ,观察pH值的改变对胚胎生长发育的影响。结果　与对照组比较 ,当培养基pH为 6 .5、6 .0和 9.0时培养胚胎的生长发育明显受到影响 (P <0 .0 5 ) ,胚胎畸形率和死亡率增高 (P <0 .0 5 )。当培养基pH为 5 .5时 ,培养胚胎死亡率达1 0 0 %。结论　全胚胎培养培养基pH条件改变 (过低或过高 )能够影响培养胚胎的生长发育。植入后全胚胎培养培养基最适pH值为 7.5～ 8.2。

Effects of Altered pH of Culture Medium on Rat Embryonic Growth and Development in Post-implantation Whole Embryo Culture

Objective To explore the optimum medium pH for post-implantation whole embryo culture (WEC). Methods Wistar rat embryos explanted gestation day 9.5 (at head fold stage) were cultured for 48 hours in rat immediately centrifugal serum(ICS) at pH 8.2 (control) or adjusted with 2N HCl or 5.6% NaHCO 3 to 8.0,7.5, 7.0, 6.5, 6.0, 5.5,8.5 and 9.0. Results At pH 6.0 there was a significant increase in the percent of anomalous (20%) and dead (60%) embryos, while at pH 5.5, none of embryos survival. Embryonic growth and development as denoted by yolk sac diameter, crown rump length, head length somites, embryo dry weight and developmental score were significantly reduced at pH 6.5, pH 6.0 and pH 9.0. Conclusions These data indicate that embryotoxicity and dysmorphology are increased with alterations of culture media pH. Medium of lower pH and higher pH can effect embryonic growth and development thus contribute to increased susceptibility to a chemical insult. The optimum medium pH for WEC is 7.5 to 8.2.