分割小鼠胚胎的简易方法和分割后二分胚的发育能力

The Simple Method for Splitting Embryo and the Survival of Demi-embryo after Bisection in Mouse

本研究改进和简化了小鼠胚胎分割方法,在普通立体显微镜下用玻璃针手工切割,每切割一枚胚胎只需1min。共切割4日龄鼠胚701枚,得1346枚可用半胚,分割成功率为96.01%。半胚培养发育率达73.70%。冷冻裸半胚,装透明带半胚和装透明带后再用琼脂包埋半胚的解冻后培养发育率分别为26.67%、56.67%和64.29%。

The objective of this study is to improve and simplify the method for splitting embryo. This study provides the simplest splitting method without micromanipulator. With the aid of a common stereomicroscope, embryos are bisected using a glass needle by hand. Only about one minute is required for each embryo to bisected in this way. A total of 701 four-day mouse embryos were bisected and 1346 demi-embryos (96.01%) were successfully produced.The developmental rate of demi-embryos in vitro culture is 73.70% ( divided blastocysts 72.88% and divided morulae 74.22% ) .There was no significant difference in the developmental rate of demi-embryos in vitro culture between blastocyst and morulae. The developmental rate of demi-embryos without zona pellucida in vitro culture of with 20% fetal calf serum or 20% sheep serum or 20% equine serum was 64.95%, 73.54% and 76.03% respectively. Moreover there was also no significant difference between them. The developmental rate of the frozen/ thawed demi-embryos with or without zona pellucida and demi-embryos with zona pellucida embedded in agar in vitro culture was 26.67%, 56.67% and 64.29% respectively.