Effect of recovery technique and culture media on in vitro maturation of indigenous cattle oocytes of Assam

Abstract

The study was undertaken to see the effect of recovery techniques and culture media on in-vitro maturation (IVM) of oocytes indigenous cattle of Assam. Oocytes were collected by aspiration and dissection techniques and were graded as A, B, C and D on the basis of their cumulus cell layers surrounding the zona pellucida. Oocytes with A, B and C categories were subjected to IVM using three different media (TCM-199, Ham's F-12 and Ham's F-10) with same additives (Sodium pyruvate, Gentamicin, L-glutamine, Porcine FSH, Follicular Fluid and HIOCS). Percentage of oocytes with expansion of cumulus cells was found to be the highest in dissection technique using TCM-199 media in comparison to Ham's F-12 and Ham's F-10 using same additives. However, it did not differ significantly between techniques. Incidence of Metaphase-I + Metaphase-II was the highest in oocytes recovered by dissection technique and cultured in TCM-199 with additives when incubated at 38.5°C for 24 hours and it differed significantly between media but there was no significant difference between the recovery techniques. In conclusion, the culture medium TCM-199 along with additives may give better IVM rate of the oocytes recovered by dissection technique from indigenous cattle ovaries of Assam.