Author:
KHARCHE S D,GOEL A K,JINDAL S K,SARASWAT SONIA,PATHAK JUHI,AGARWAL SURBHI
Abstract
The present study was undertaken to investigate the effect of granulosa cell co-culture, granulosa monolayer and hormone on in vitro maturation of nude caprine oocytes. Nude oocytes (644) having evenly granulated cytoplasm were recovered by puncture technique. The oocytes were divided into 6 groups and matured in TCM-199 medium supplemented with 10% fetal bovine serum (FBS) and granulosa cell co-culture/ granulosa monolayer/ hormones. After maturation (27h), the oocytes were fixed for 15 min in 2.5% (w/v) glutaraldehyde in phosphate buffer saline,washed twice with PBS, stained with 0.1mg/ml DAPI in PBS and mounted on slides. The evaluation of nuclear status was done by epiflourescence microscope by observing either a polar body release or presence of two shiny chromatin spots. The maturation rate of oocytes in group 1, 2, 3, 4, 5 and 6 were 23.47, 32.40, 31.77, 26.00, 38.09 and 51.37%, respectively. The results of the study indicated that TCM-199 supplemented with hormones and coculture granulosa cells significantly increases the maturation rate of in vitro matured nude caprine oocytes.
Publisher
Indian Council of Agricultural Research, Directorate of Knowledge Management in Agriculture
Subject
General Veterinary,Animal Science and Zoology