Effects Of Different Culture Medium Compositions And Culture Systems On Buffalo Embryo Development In Vitro

The objective of the present study is to assess the effect of different culture systems and culture medium compositions on in vitro early embryo development in buffalo and, based on this study, to subsequently optimise the culture system for a better in vitro early embryo development in buffalo.Oocytes were aspirated from 2-6 mm follicles on the ovaries from slaughterhouse and matured in vitro.After maturation,oocytes were fertilized with buffalo sperm . 18h after insemination, the presumptive zygotes were transfer to culture medium supplemented with different compositions.Cleavage and blastocyst rates were recorded on D2 and D5~D9 after insemination, respectively.Cells of each blastocyst were counted with fluorescent microscope after staining with Hoechst33342. Hatching of embryos post frozen-thawed were assessed.Analysis of different protein expression in embryos was carried out with SDS-PAGE electrophoresis.The results showed that: i) The cleavage rate,blastocyst rate,cell numbers of blastocyst and the hatching rate post frozen-thawed were significantly higher than other groups (P<0.05), when the concentration of insulin in the culture medium was 5 μg/mL; ii) There was a negative effect on early embryo development and the hatching rate,when the supplementation of glucose was higher than 6 111M in the embryo culture medium at 18 h or 96 h after insemination; iii) A combination of supplementations with insulin and glucose in the culture medium could significantly increase the blastocyst development rate,cell numbers in blastocyst and hatching rate(P<0.05); iv) Both supplementations of EAA at 10 h and NEAA at 96 h in the culture mediun after insemination could increase the developmental potential (P< 0.05); v) Supplementation of sodium citrate and myo-inositol had no significant effect on the buffalo embryo development as comparing the controls (P>0.05). vi) The cell...