Effect Of Cumuus Cells On In Vitro Maturation Of Buffalo Oocytes
By: Hassan Mahmood Warriach
| Dr.Kazim Raza Chohan.
Contributor(s): Dr.Rashid Ahmad Chaudhry | Faculty of Veterinary Sciences.
Material type: 
BookPublisher: 2001Subject(s): Department of TheriogenologyDDC classification: 0718,T Dissertation note: In present study, the meiotic competence of buffalo oocytes with different number of cumulus cell layers was compared. A total of 775 ovaries in 9-replicates were collected from Nili-Ravi buffaloes from a local abattoir. A total of 441 oocytes were aspirated with a disposable syringe using 18 gauge needle and an average of 0.78 oocytes were collected per ovary. Oocytes were divided into four groups as, oocytes with 3-5 layers of cumulus cells, oocytes with 1-2 layers of cumulus cells, denuded oocytes and oocytes to be co-cultured with cumulus cells. Oocytes in all four groups were matured in mediurn-199 with Earle's salts supplemented with 100 rng/Lglutamine, 2.2 g/L NaHCO3, 10ug/mI streptornycin, 100 IU/mi penicillin, 10 mM HEPES and 10% fetal calf serum. Around 100 oocytes in each group from 9 replicates were transferred to 100 u1 droplets (5 to 8 oocytes/drop) of maturation medium under sterile mineral oil in 60 x 35 mm sterile plastic dishes for 24 hours at 39°C and 5% CO2. After 24 hours of culture, cumulus cells were removed from oocytes. Denuded oocytes were fixed, stained with DAPI and evaluated for rneiotic competence (oocytes reaching to metaphase-Il) under fluorescent microscope. The oocytes with 3-5 layers of cumulus cells showed significantly (P < 0.05) higher maturation rates (64.5%) than denuded (8.6%) and co-cultured oocytes (34.5%) but did not differ (P> 0.05) from oocytes having 1-2 layers of cumulus cells (51.4%). The degenerate rates were significantly higher (P <0.05) for denuded oocytes (51%) than all other groups (Average: 13.8% to 17.4%). In conclusion the results of present study suggested that cumulus cells have positive influence on oocyte maturation and co-culture of denuded oocytes with cumulus cells improved meiotic competence.
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UVAS Library Thesis Section | Veterinary Science | 0718,T (Browse shelf) | Available | 0718,T |
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In present study, the meiotic competence of buffalo oocytes with different number of cumulus cell layers was compared. A total of 775 ovaries in 9-replicates were collected from Nili-Ravi buffaloes from a local abattoir. A total of 441 oocytes were aspirated with a disposable syringe using 18 gauge needle and an average of 0.78 oocytes were collected per ovary. Oocytes were divided into four groups as, oocytes with 3-5 layers of cumulus cells, oocytes with 1-2 layers of cumulus cells, denuded oocytes and oocytes to be co-cultured with cumulus cells. Oocytes in all four groups were matured in mediurn-199 with Earle's salts supplemented with 100 rng/Lglutamine, 2.2 g/L NaHCO3, 10ug/mI streptornycin, 100 IU/mi penicillin, 10 mM HEPES and 10% fetal calf serum. Around 100 oocytes in each group from 9 replicates were transferred to 100 u1 droplets (5 to 8 oocytes/drop) of maturation medium under sterile mineral oil in 60 x 35 mm sterile plastic dishes for 24 hours at 39°C and 5% CO2. After 24 hours of culture, cumulus cells were removed from oocytes. Denuded oocytes were fixed, stained with DAPI and evaluated for rneiotic competence (oocytes reaching to metaphase-Il) under fluorescent microscope. The oocytes with 3-5 layers of cumulus cells showed significantly (P < 0.05) higher maturation rates (64.5%) than denuded (8.6%) and co-cultured oocytes (34.5%) but did not differ (P> 0.05) from oocytes having 1-2 layers of cumulus cells (51.4%). The degenerate rates were significantly higher (P <0.05) for denuded oocytes (51%) than all other groups (Average: 13.8% to 17.4%). In conclusion the results of present study suggested that cumulus cells have positive influence on oocyte maturation and co-culture of denuded oocytes with cumulus cells improved meiotic competence.
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