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|Title:||Development of in vitro embryo production systems for red deer (Cervus elaphus) - Part 2. The timing of in vitro nuclear ooctye maturation|
|Citation:||Animal Reproduction Science, 2002; 70(1-2):77-84|
|Publisher:||Elsevier Science BV|
|D. K. Berg, J. G. Thompson, and G. W. Asher|
|Abstract:||The time course of in vitro red deer nuclear oocyte maturation was determined. Ovaries were obtained at slaughter and oocytes were aspirated from follicles greater than 2 mm in diameter. Oocytes with compact cumulus cells were matured in 50 μl microdrops (10 per drop) under mineral oil containing TCM 199 supplemented with 0.33 mM pyruvate, 10 μg LH and FSH, 1 μg oestradiol and 10% foetal bovine serum. Oocytes were matured at 39 °C and 5% CO2 in air. At 3 h intervals (0–27 h) oocytes were removed from incubation, cumulus expansion scored and removed, and fixed oocytes in ethanol:acetic acid (3:1) for 48 h. Oocytes were stained with lacmoid (1%) and nuclear maturation assessed. Oocytes were arrested in the germinal vesicle (GV) stage at aspiration and up to 6 h of incubation. The nuclear membrane began to disperse after 6 h and by 10.6±0.6 h of incubation 75% of the oocytes exhibited germinal vesicle breakdown (GVBD). The mean time for 50% of the oocytes to reach metaphase one (MI) and metaphase two (MII) was 11.7±0.4 and 24.8±0.9 h, respectively. Cumulus oophorus were tightly compacted at aspiration and did not begin expansion until 12 h of culture. Full expansion was complete by 18 h of culture. Corona radiata cells did not begin expansion until 15 h and were fully expanded by 24 h. Results indicate that in vitro red deer oocyte maturation follows a similar time course of nuclear maturation as reported for bovine and ovine oocytes.|
|Keywords:||Red deer; Cervus elaphus; In vitro oocyte maturation|
|Description:||Copyright © 2002 Published by Elsevier Science B.V. All rights reserved.|
|Appears in Collections:||Obstetrics and Gynaecology publications|
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