小鼠电融合胚胎发育影响因素的研究

电融合技术，是获得四倍体胚胎的主要手段．本试验研究了各种不同因素，对小鼠电融合胚发育的影响．结果表明，电刺激前后，使用不同的操作液对融合胚发育有影响．电刺激前使用Ｍ２液、电刺激后在Ｗｈｉｔｔｅｎ氏液中融合，然后转入ＣＺＢ液中培养，这一操作体系（Ｍ／Ｗ—ＣＺＢ）效果最好，融合胚的囊胚发育率为８０．５％，显著高于Ｍ／Ｗ—Ｗ，Ｗ／Ｗ—Ｗ，Ｗ／Ｗ—ＣＺＢ组，后者的囊胚发育率分别为０，０，５４．５％（Ｐ＜０．０５）．试验还发现，刺激强度为１．０ｋｖ／ｃｍ，８０μｓ；１．０ｋｖ／ｃｍ．４０μｓ和０．８ｋｙ／ｃｍ，８０μｓ时，融合囊胚的发育率分别为７９．０，８１．９和７１．１％．ＨＣＧ注射后４７ｈ、４８ｈ的２—细胞胚胎的融合胚发育率，显著高于（Ｐ＜０．０１）ＨＣＧ注射后４２ｈ、４４ｈ和４６ｈ的，囊胚发育率分别为８２．９，８０．６，０，０和２２．８％．

FACTORS AFFECTING THE DEVELOPMENT OF ELECTROFUSED MOUSE EMBRYOS

Electrofusion of 2-cell embryos is the major method for the production of tetraploid embryos. In order to establish a appropriate system to obtain more tetraploid blastocysts, we studied several factors affecting the development of fused mouse embryos. Percentage of blastocysts developing from fused embryos was higher (p<0.05) when the working media used was M2 before and Whitten medium after stimulation, and the culture medium was CZB (M/W-CZB)(80.5%), than that from other combinations or media (M/W-W, 0%;W/W-W, % and W/W-CZB, 54.5%). The rates of blastoeysts were significantly higher when electrie stimulus were 1.0kv /cm, 40 as(81.9%); 1.0kv / cm,80μs (79.0%); 0.8 kv.cm,80μs (71.l%) than that with 1.0 kv/cm, 20μs (55.2%); 0.8 kv/ cm lasting 20μs (25.7%);40μs (38.3%),and 160μs (27.3%), respectively. Significant difference was also observed in fused blastocyst rate when the 2-cell embryos were obtained 47 h (82.9%) and 48 h (80.6%) after the HCG-injection and when the 2-cell embryos were collected 42 h (0%) and 44 h (0%),and 46 h (22.8%) after HCG-injection, respectively.