An efficient method for in vitro fertilization in rabbits

This experiment was carried out to study a simple and efficient method for in vitro production of rabbit embryos. Newly ejaculated rabbit spermatozoa were used to fertilize superovulated oocytes after capacitation in vitro with four different media: (A) isotonic defined medium (DM)+heparin, (B) DM only, (C) DM+ high ionic strength defined medium (HIS), and (D) DM supplemented with 10mM NaHCO3 (mDM) +HIS supplemented with 10mM NaHCO3 (mHIS). The presumptive zygotes were cultured in M199 supplemented with 10% FCS, 1.25mM Na Pyruvate and 0.1mM EDTA (mM199). The cleavage rates after 24h of incubation were 29.3%, 32.1%, 64.9%, and 91.6% respectively, and the rates of blastocyst formation after 72h were 0, 27.3%, 58.4% and 85.2%, respectively. The results in the (D) treatment were significantly better than the other three treatments (p<0.01). Developmental potential of in vivo and in vitro derived zygotes was also compared using the mM199. The percentages of blastocyst and hatching blastocyst in the two groups were 92.5% and 87.2% after 84h, and 84.9% and 83.7% after 108h, respectively, and the two groups were not significantly different (p>0.05). The developmental progress of the two groups was nearly synchronous towards the end of culture. When IVF embryos from 2- to 4-cell stage were transferred into recipients, the pregnancy rate did not differ from in vivo fertilization, but the rate of live young from IVF was significantly lower than from in vivo. The results of this experiment showed that ejaculated rabbit sperm could be capacitated efficiently after treatment of mDM and mHIS, and rabbit NF embryos achieved great development in mM199 in vitro.