Different Culture Systems Yanbian Cattle Somatic Cell Cloned Embryos

Because of the specious character of Yanbian Yellow Cattle in meat, it is famous inthe international market, but it has some shortcomings, such as, growth rates slow, theperformance of product meat low, the coverage rate of eminent strain low and so on, thedominant courses are attributed to the development of technology, system and cost. Theefficiency is lower in improving and selection, breeding periods is longer. Recently withthe development of somatic cell cloning, it has made breeding periods shorten severaltens of years, comparing with conventional breeding and improving methods. If it cancooperate with past breeding technology system, it can improve the breeding potentialgreatly in eminent strain, it is important to accelerate resource protection of YanbianYellow Cattle, and then it has a great importance in Yellow Cattle product.In this research, we use fresh granulosa cells as donor cells; we use MⅡenucleatedoocytes as recipient cells. We do the experiment by means of acupressure enucleatedoocytes putting donor granulosa cells into yolk space, with 0.3M mannitol fusion, 20μsone electric fused push, 5μM Ionomycin treating 5min, 2mM 6-DMAP activation 4h, according to experiment designing, put them into different cultures, change culture fluidregularly and observe the embryo developmental conditions, the result is evaluated ascleavage rate, 4-6 cell developmental rate and 8-16 cell developmental rate. They areshown that:(1) Culture in CR1aa and TCM-199 culture medium, 4-6 cell developmental rate and8-16 cell developmental rate is 63.6％, 57.9％and 27.1％, 28.2％, respectively. It issignificantly higher than mSOF culture medium 43.2％and 17.3％.(2) Adding 1mM GSH in CR1aa culture medium, cleavage rate, 4-6 celldevelopmental rate and 8-16 cell developmental rate is 61.0％, 66.0％and 45.2％, respectively. It is superior to adding 2mM and 3mM treatment groups.(3) Without adding serum before 72h, 4-6 cell developmental rate is higher thancontrol group CR1aa (69.8％vs 55.6％), but 8-16 cell developmental rate is lowerthan control group (19.9％vs 31.0％), there is significant difference in them(P＜0.05). Adding glucose at the fifth day to cultural medium, 8-16 celldevelopmental rate is prominently increased (40.5 vs 31.0％).(4) Adding ouiduet epithelial cell or granulose cell to CR1aa or TCM-199 co-culture, the cleavage rate, 4-6 cell developmental rate and 8-16 cell developmental aresignificantly increased (P＜0.05).We can conclude that: CR1aa and TCM-199 containing 5％FBS were the samewith somatic cell cloning in culture in vitro in Yanbian Yellow Cattle. Adding 1mMGSH to CR1aa containing 5％FBS, the developmental rate in vitro can be greatlyincreased in somatic cell cloning in Yanbian Yellow Cattle. Adding serum to the culturemedium in the former period culture in vitro, the developmental ability can be restrained, and then adding glucose to culture medium in the latter period culture in vitro, it can beimproved. Adding ouiduct epithelial cell or granulose cell to CR1aa or TCM-199co-culture, the developmental abilities can be significantly increased.