包被方法对小鼠半胚冷冻效果的影响

将小鼠囊胚一分为二后,采用血清-琼脂法(A),透明带-琼脂法 (B)和双层透明带法(C)包被半胚。在RPE冷冻仪的控制下,对包被半胚进行冷冻。解冻后将360枚半胚进行体外培养。A,B,C三组的半胚发育率分别为69.2％(83/120),67.5％(81/120)和60％(72/120)。将12对在冷冻前用A法包被的解冻半胚移植于6只假孕母鼠,结果有3只妊娠,共产半胚鼠8只,其中3对为同卵双生。实验证明,本研究建立的血清-琼脂包被法操作简便,效果良好,便于应用,是一种较理想的包被半胚新方法。

Effect of Embedmental Methods on Freezing of Demi-Embryos in Mouse

Blastoeysts of mouse were bisected and demi-embryos were embedded by following methods:(A)demi-embryos without Zona Pellucida (ZP) were embedded with serum and agar,(B)demi-embryos insorted,into ZP were embedded with agar,(C)demi-embryos with ZP were inserted into an additional ZP.The embedded demi-embryos were frozen.And after thawing,360 demi-embryos were cultured in vitro.The proportion of demi- embryos that developed into blastoeysts in group A,group B and group C was 69.2%(83/120),67.5%(81/220) and 60%(72/220) respectively.Twelve pairs of forzen-thawed demi-embryos that were not cultured after removal of glycerol were trarsferred into 6 pseudopregnantrecipients,3 of which become pregnant,and 8 young mice were obtained,including 3 pairs of mono- zygotic twins.The present study demonstrated that high survival of frozne- thawed demi-embryos can be obtained by embedding demi-embryos without ZP with serm and agar before freezing.