Preliminary Studies On Intra-and Inter-Species Fertilization Of Porcine Oocytes Subjected To Intracytoplasmic Sperm Injection

This thesis was designed to study the issues related to intra- and inter-species fertilization of porcine oocytes subjected to intracytoplasmic sperm injection.The text was arranged into four parts:the first part included(1) Effect of supplementation method of follicile stimulating hormone(FSH)+ luteinizing hormone(LH)during porcine oocyte in vitro maturation on futher development after parthenogenetic activation;(2)Effect of supplementation of different cysteine concentrations during porcine oocyte in vitro maturation on futher development after parthenogenetic activation;(3)Effect of supplementation of different epidermal growth factor(EGF)concentrations during porcine oocyte in vitro maturation on further development after parthenogenetic activation.The second part included the effect of different culture medium volume on early development of porcine parthenogenetic embryos.The third and fouth parts included(1)Effect of different activation methods on pronuclear formation and sperm decondensation of porcine oocytes after intracytoplasmic sperm injection(ICSI);(2)Effect of different activation methods on early development of porcine oocytes after ICSI;(3)Effect of injection of cynomolgus monkey and buffalo spermatozoa into porcine oocytes on the pronuclear formation after ICSI;(4)Effect of cynomolgus monkey and buffalo spermatozoa into porcine oocytes on early development of porcine ICSI embryos.It was concluded that:in the first part(1)the way of adding follicile FSH + LH for 22h,and changing new maturation with FSH + LH afer 22h maturation in vitro;(2)adding 1.2 mmol/L cysteine in maturation medium;(3)adding 10 ng/ml EGF in maturation medium,were suitable for the in vitro maturation of porcine oocytes and further parthenogenetic development.In the second part the average ratio of embryo/volume being 1:2-3μL is suitable for porcine parthenogenetic embryo in vitro development.In the third and fouth parts(1) there was no significant difference on pronuclear formation and blastocyst formation of porcine ICSI oocyte between the two different activation treatments;(2)Pronuclear formation and blastocyt formation were observed in the porcine oocytes injected with cynomolgus monkey spermatozoa and buffalo spermatozoa separately.