The Study Of Parthennogenetic Activation And Embryo Culture Of Porcine Oocytes And The Effects Of Cell Factors On The Maturation Of Porcine Oocytes

The culture of porcine oocytes maturation in vitro, parthenogenetic activation of MII porcine oocytes and embryo culture are main technology of embryo biology engineering. People gradually focus on the elementary technology of porcine oocytes development and its mechanism. Optimizing culture conditions of porcine oocytes maturation in vitro, parthennogenetic activation of MII porcine oocytes and embryo culture and clarifying its mechanism are important to somatic cell nuclear transfer (SCNT) technology and transgenic animal production. Nowadays many studies have been carried out in the above three aspects in the foreign country and home. But the studies aren't still mature and affect our progress of somatic cell nuclear transfer and the study of its mechanism. So our experiments study the culture of porcine oocytes maturation in vitro, parthenogenesis of MII porcine oocytes, embryo culture, the effects of IGF-I, bFGF, TGF-α separation or combination on porcine oocytes maturation in vitro and oocytes cleavage after parthennogenetic activation and the mechanism of 1GF-I promoting porcine oocytes maturation in vitro in order to establish the technology of porcine oocytes development in vitro and apply the technology to the study of transgenic animal and mechanism of somatic cell nuclear transfer.Our study includes four aspects. In the first aspect we study several important conditions of porcine oocytes maturation in vitro and oocytes cleavage after parthenogenetic activation and found mNCSU-23+15IU/mlPMSG+20IU/mlHCG+15% PFF+0.57mMcysteine is a good culture condition .When the Cocs are cultured in it ,the maturation rate and oocytes cleavage rate are higher than those of foreign covered. Our result are (86.7±3.35)% and (86.3±4.16)% and the highest report of foreign is(85.7±4.1)%.In the second aspect we study the effect of different chemical activations on development of porcine parthennogenetic embryo and found two best activation method. The first one is that putting the maturation MII oocytes in the 20μmol/L ionomycin for 30 minutes and then putting them in the NCSU-23 condition containing 5μg/mICB and 5mM/L6-DMAP for 3.5 hours, the oocytes cleavage rate and morulae/blastocysts development rate are (76.7±7.6)% and (37.1±6.4)%.The second one is that putting the maturation MII oocytes in the 200μM/L Thimerosal for 20 minutes and then putting them in the NCSU-23 condition containing 8mM DTT for 30 minutes,...