In vitro maturation of nude caprine oocytes recovered during oocyte collection
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Authors
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S D KHARCHE
Principal Scientist, Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281122 India
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A K GOEL
Principal Scientist, Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281122 India
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S K JINDAL
Principal Scientist, Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281122 India
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SONIA SARASWAT
Research Associate, Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281122 India
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JUHI PATHAK
Senior Research Fellow, Physiology, Reproduction and Shelter Management Division, Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281122 India
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SURBHI AGARWAL
Senior Research Fellow, Physiology, Reproduction and Shelter Management Division, Central Institute for Research on Goats, Makhdoom, Uttar Pradesh 281122 India
Keywords:
Caprine, In vitro maturation, Nude oocytesAbstract
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.
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