22-Title: Effect of growth factors on in vitro maturation and in vitro culture of buffalo oocytes
Authors: Vijay Singh, AK Misra, Suresh Kumar and Vijay Kumar
Source: Ruminant Science (2015)-4(2):219-224.
How to cite this manuscript: Singh Vijay, Misra AK, Kumar Suresh and Kumar Vijay (2015). Effect of growth factors on in vitro maturation and in vitro culture of buffalo oocytes. Ruminant Science 4(2):219-224.
The objective of this work was to study the effect of growth factors (EGF and IGF-I) on in vitro maturation, fertilization and development of buffalo (Bubalus sp.) cumulus oocyte complexes (COCs). Buffalo ovaries were collected from abattoir and transported to laboratory within 2 h of slaughter. Oocytes aspirated from 3-10mm follicles through 18 G needle using 10 ml syringe in oocyte collection medium, were classified as A, B, C and D grades (depending upon the cumulus cells attached and even granulation) and only grades A, B and C were used for in vitro maturation. The oocytes were matured in two groups [six treatment groups comprised of 10, 20, 30 ng/ml of EGF or 10, 50 or 100 ng/ml of IGF-I in the IVM media whereas seventh group (without EGF /IGF-I) served as control]. Matured oocytes were co-incubated with ~ 1×106/ml sperms of Murrah bull in mSOF medium with heparin (10µg/ml) and after 22 h of oocyte-sperm incubation, fertilized oocytes were stripped of cumulus cells and cultured in mSOF medium for 8 days to study embryo development. Oocytes maturation rate improved significantly (P<0.05) at all concentrations of EGF or IGF-I compared to control. Following IVF, cleavage was significantly (P<0.05) high at 20 or 30 ng/ml EGF/50 or 100 ng/ml IGF-I, compared to control .Development of embryos to morula stage was significantly (P<0.05) improved with 20 ng/ml EGF or all the three concentrations of IGF-I. Significantly (P<0.05) more blastocyst developed at 100 ng/ml of IGF-I. It was concluded that supplementation of IVM media with either EGF or IGF-1 significantly improves the in vitro maturation and subsequent development of buffalo oocytes.
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