Veterinary Archives, Vol. 84 No. 5, 2014.
Original scientific paper
Effect of growth factor and antioxidant on in vitro maturation of oocytes and cleavage rates of in vitro produced Indian buffalo (Bubalus bubalis) embryos.
Enchaparambil M. Sadeesh
; Animal Physiology and Reproduction Division, ICAR-Central Institute for Research on Buffaloes, Hisar, India
Fozia Shah
; Department of Veterinary Physiology and Biochemistry, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, India
Ashok K. Balhara
; Animal Physiology and Reproduction Division, ICAR-Central Institute for Research on Buffaloes, Hisar, India
Sumangali M. K. Thirumaran
; Animal Genetics, Breeding and Management Division, ICAR-Central Institute for Research on Buffaloes, Hisar, India
Sarita Yadav
; Animal Genetics, Breeding and Management Division, ICAR-Central Institute for Research on Buffaloes, Hisar, India
Prem S. Yadav
; Animal Physiology and Reproduction Division, ICAR-Central Institute for Research on Buffaloes, Hisar, India
Abstract
The present study was carried out to evaluate the effect of growth factor in combination with antioxidant on the in vitro maturation (IVM) rate of oocytes and cleavage rates of in vitro fertilized (IVF) Indian Bubalus bubalis embryos. The cumulus oocyte complexes (COCs) were collected from slaughterhouse ovaries by aspirating follicles, and kept in maturation media (MM) for 24 h. The MM consisted of Group-1 (Control MM) - TCM 199 + 10% FBS + PMSG (20 IU/mL) + hCG (10 IU/mL) + Sodium pyruvate (0.80 mM) + L-glutamine (2 mM) + Gentamicin (50 μg/mL), Group-2 - Control MM + Epidermal growth factor (EGF) (10 ng/mL) + β-mercaptoethanol (β-ME) (25 μM), Group -3 - Control MM + EGF (20 ng/mL) + β-ME (100 μM), Group-4 - Control MM + EGF (100 ng/mL) + β-ME (500 μM). After maturation in diverse maturation media, the media used for IVF and the subsequent development of embryos was the same in all groups. The present results revealed that 20 ng/mL EGF + 100 μM β-ME concentrations were optimum and showed a significant effect on oocyte maturation and further development. Also high concentrations of EGF + β-ME (100 ng/mL + 500 μM), showed a decrease in the cumulus expansion rate, polar body formation rate and cleavage rates. A significant improvement in cleavage rate was observed when oocytes were matured in maturation medium with 20 ng/mL EGF + 100 μM β-ME and capacitation and fertilization was carried out in BO medium compared with TALP medium (64.8 ± 3.8 vs 44.1 ± 2.9). No significant difference in cleavage rate was observed for oocytes matured in the control group, control group + 10 ng/mL EGF + 25 μM β-ME and control group + 100 ng/mL EGF + 500 μM β-ME, between BO and TALP medium, indicating the beneficial effect of the addition of 20 ng/mL EGF + 100 μM β-ME to the maturation media for improved Bubalus bubalis embryo production under in vitro culture.
Keywords
oocyte; growth factor; antioxidant; in vitro maturation; in vitro fertilization; cleavage rate
Hrčak ID:
128406
URI
Publication date:
10.10.2014.
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