Studies On Semen Cryopreservation And In Vitro Fertilization Of Swine

This experiment was conducted to study techniques of Semen Cryopreservation andtechniques of oocytes in vitro maturation (IVM), in vitro fertilization (IVF) and development ofearly embryos, as to enhance the efficiency of both Semen Cryopreservation and in vitrocultivation system.1．Study on WZSP Semen Cryopreservation TechniquesIn this experiment, semen pellets were frozen on the fluorin plant. Sperm motility, viabilityand acrosome integrity were studied in this experiment to select better freezing extenders,balancing hours, semen plasma remnants and freezing-thawing protocols, the results were asfollows, (1) on semen cryopreservation, effects of both No.2 and No.3 extenders with amino acidin them were better significantly than that of No.1 extender with saccharide in it only(P<0．05) ;both the sperm motility and viability treated with No.4 extender (with 2．5ml DMSO) after thawingwere significantly lower than those of sperms treated with No.2(with 2ml glycerin) and No.3(with1．25 ml DMSO&1ml glycerin) extenders(P<0．05) . (2) both the thawing sperm viability andplasma acrosome integrity of semen treated with 4℃for 2h were better than those at 17℃C for 3hand then at 4℃for 2h before frozen, and the sperm motility had significant differences(P<0．05) (3) both sperm motility and viability of the group remaining 0ml/10ml semen plasma remnants werelower significantly than those of the other 2 groups remaining 0．5ml/10ml or 1ml/10ml semenplasma remnants (P<0．05) . (4) the effects of dried thawing were better than those of thawing insolution significantly (P<0．05) .2．Study on in vitro Fertilization and development of early embryosThis experiment was about the effects of different culture mediums, different sperms originand input of Traditional Chinese Medicines on oocytes IVM and early growth of IVF embryos. Itsuggested that, (1) the rate of oocytes maturation was significant higher after cultured inNCSU-23(75．45%) than in TCM-199(64．38%)(P<0．05) ; but effects of both NCSU-23(10．24%)and TCM-199(9．42%) were similar on development of early embryos (P>0．05) . (2) the cleavagerate(67．96%) and morula rate(44．66%) of embryos developed from thawing semen were similar tothose from fresh semen(67．41% & 45．09%)(P>0．05) ; the blatocyst rate(10．27%) of embryosdeveloped from fresh semen was significantly higher than that from frozen semen(4．37%)(P<0．05) . (3) the embryos developed from fresh sperms were cultured in mediumcontaining 0．10%(V/V) Cistanche or 0．10%(V/V) Radix Astragali; blatocyst rate of the embryoscultured in medium containing Cistanche (15．76%) was higer than that of embryos cultured inmedium without Traditional Chinese Medicine markedly(9．26%)(P<0．05) , and it was also a littlehigher than that of embryos cultured in medium containing Radix Astragali (10．09%)(P>0．05) . (4) the blatocyst rate of embryos cultured in the medium containing 0．05% and the mediumcontaining 0．10%(V/V) Cistanche(17．02% & 16．75%) were higher notablely than those ofembryos cultured in the medium containing 0．15%(V/V) Cistanche (0%)(P<0．01) or the mediumcontaining no Cistanche(8．94%)(P<0．05) . (5) the blatocyst rate of embryos transferred into themedium containing 0．10%(V/V) Cistanche at 2-cells time (13．81%) was significantly higher thanthat at 4-cells time (7．53%)(P<0．05) ; it was also a litter higher than that at oosperm time (13．67%)(P>0．05) .