In Vitro Maturation Of Oocytes From Slaughtered Goats

In recent years, the business that embryo transplant application and the research of every kind of embryo engineering development all need a mass of high-quality earlier period embryo. The source and the cost of the embryo has become the important factor that practical turn of embryo engineering. Making use of the oocytes from Slaughtered Goats FVM (in vitro maturation), joining together the Bore goat sperm capacitation and the embryo transplants, we could investigate the low cost technique route that improves the goat species of our country.Ovaria from slaughtered goats were carried to a laboratory at 37.0℃~39.0℃ in 2-3 hour. From an ovary could be obtained 1.43 Cumulus oocytes complexes (COCs). The result showed that slicing obtained significantly more useable oocytes than aspiration.There were 365 oocytes collected from the Graafian follicles (2~6mm in diameter) of newly slaughtered goat have been grouped into three type: Type A, the oocytes surrounded by cumulus cells; Type B, the oocytes surrounded by partly by cumulus cells; Type C, denuded oocytes. Goat oocytes of grade A& B are selected for in vitro maturation (FVM).These oocytes have been cultured in three different kinds of medium. 1), Ml99 + Pencillin(75^g/ml)+Streptomycin(60ug/ml)+10%FBS+LH(20μg/ml)+FSH(10IU/ml);2),M 199+Pellcillin(75μg/ml)+Streptomycin(60μg/ml)+15%FBS+LH(20μg/ml)+FSH(10IU/ml); 3),M199+10%FBS+LH(20ng/ml)+FSH(10IU/ml),at 38.0℃~39.0℃under water- satured atmosphere of 95% air-5% CO2 for 24~26 hour. The percentage of oocytes maturation canbe calculated under the microscope. Out of a total number of 365 cultured oocytes, 301(82.5%) matured oocytes were obtained. The results suggested that an addition of 10%~15%FBS, 10IU/ml FSH, and 20ug/ml LH was effective to improve oocyte maturation, and to extrude the first polar body.In the present experiments, the function of NCS on goat oocytes for IVM was studied. The results were as follows: the in vitro maturation rate of the follicular oocytes in the medium with FSH and LH supplemented with 10% NCS (83.8%) was higher than that of with 10%FBS (81.7%, P>0.05). This research established groundwork for investigating suitable mature system of the oocyte IVM for actual production demand.Follicular oocytes were collected from new slaughtered goat's ovaries. 345 cumulus-oocyte combinations (COCs) of grade A &B were cultured respectively in the mediums of M199+Pencillin(75μg/inl)+Streptomycin(60μg/inl)+LH(20μg/ml)+FSH(10IU/ml),M199+ 10%FBS+Pencillin(75μg/ml)+Streptomycin(60μg/ml>l-LH(20μg/ml)+FSH (10 IU/ml), M199+15%FBS+Pencillin(75μg/ml)+Streptomycin(60μg/ml)+LH(20μg/ml)+FSH(10IU/ ml).The matur -ation rates were 0%, 81.7% and 81.7% respectively. It is concluded that 10%~15% FBS is beneficial to the maturation in vitro of goat follicular oocytes.In this experiment, the effects of temperature on goat oocytes IVM were studied. The results suggested that the effects of the temperature of 38.0℃ > 38.5℃ and 39.0℃ were not different on the IVM rate of the follicular oocytes (P>0.05). The effects of culture time on goat oocytes IVM were studied. The maturation rate of oocytes cultured for 24,26h was significantly higher than that of 16h (P<0.05).In the present experiments, the ability of bacteriophage on goat oocytes FVM was studied.The results suggested that an addition of 75ug/ml Pencillin+60ug/ml Streptomycin reduce the oocytes maturation rate. But the effect was not prominent (P<0.05). This result indicated that the Goat oocytes IVM is could put in practice for actual production.