The Study On Superovulation And In Vitro Embryo Production In Lambs

The northeast fine-wool sheep was a fine varieties used for meat and wool dual-purpose, this study investigated young female lambs’ superovulation, oocyte in vitro maturation, in vitro fertilization and embryo culture, this technology is designed to provide practical applications in production reference.The experience included three parts: the factors of lamb superovulation, the effects of five oocytes maturation solution and fertilized egg influencing factors. The results as follows:1. The recovered eggs, and eggs of 6-week-old lamb was higher than 4-week-old group and 8-week-old group, but the difference was not significant(P>0.05), the available egg rate was significantly higher than 4-week-old group(P <0.05). The recoveries and available oocytes rate which used by FSH+PMSG(produced in Canada) was higher than the other three groups, and was significantly higher than FSH+LH(domestic) group(P<0.01). The available oocyte rate in autumn was higher than in the spring and was significantly higher than summer(P<0.01). The recoveries of superovulation of the first time was significantly higher than repeat superovulation group(P<0.01), The available oocytes of non-transportation group was significantly higher than the rate of transport group(P <0.01), and the recoveries was no significant difference(P>0.05).2. This experiment compared the effect of five oocyte mature culture solution, the maturation rate among each group was not significant(P>0.05), it showed that 10mg/ml BSA could replace serum(10% FBS) added.3. In the same capacitation time, the control group was significantly lower than capacitation heparin added group(P <0.05), the capacitation rate of heparin added group reached a peak after 2 hours, and was significantly higher than control group(P<0.05). 25μg/mL heparin added group sperm survival time was significantly lower than the control group(P<0.05), while it was significantly higher than 50μg/mL, 100μg/mL added group(P<0.05); Consolidated results showed that use 25μg/mL heparin concentration, for 2 hours works best. The PBII rate of oocyte diameter greater than 130μm group was significantly higher than that of d<120μm group, and cleavage rate was significantly greater than 120μm<d<130μm group(P<0.05) and significantly higher than d<120μm group(P<0.01). The PBII rate and cleavage rate of oocyte maturated for 24~27 hours was higher than that of 27~30 hours, and significantly higher than cultured 21~24 hours(P<0.01). The EGF added group could significantly increase the rate of PBI rate and cleavage rate(P<0.05), significantly increased the rate of blastocysts(P<0.01). The blastocyst rate of 20 ng/mL dosage was significant increased compared with 10 ng/m L added(P <0.01).SOF fertilization system cleavage rate was significantly higher than TALP fertilization system(P<0.05), while the SOF system blastocyst rate was significantly higher than TALP fertilization system(P<0.01).