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Development of prepubertal goat oocytes after their in vitro maturation and chemical activation

Published online by Cambridge University Press:  10 August 2020

Seungbum Hong
Affiliation:
Reproductive Biotechnology Centre, Post Box 299003, Dubai, United Arab Emirates
Binoy S. Vettical
Affiliation:
Reproductive Biotechnology Centre, Post Box 299003, Dubai, United Arab Emirates
Nisar Ahmad Wani*
Affiliation:
Reproductive Biotechnology Centre, Post Box 299003, Dubai, United Arab Emirates
*
Author for correspondence: Nisar Ahmad Wani. Reproductive Biotechnology Centre, Post Box 299003, Dubai, United Arab Emirates. Tel: +971 4 327 2568. Fax: + 971 4 327 2563. E-mail: [email protected]

Summary

Experiments were conducted to study in vitro maturation of prepubertal goat oocytes and their developmental potential after chemical activation. In Experiment 1, cumulus–oocytes complexes collected from the ovaries of prepubertal goats slaughtered at a local abattoir were matured in vitro in TCM-199-based medium supplemented with 10 µg/ml luteinizing hormone (LH) (treatment 1) or 10 µg/ml LH + 0.1 mM l-cysteine (treatment 2). In Experiment 2, mature oocytes were activated with either 5 µM ionomycin or 7% ethanol. After 18 h, some oocytes were randomly fixed and stained to evaluate their chromatin status, while others were cultured in embryo culture medium to study their further development. In Experiment 3, oocytes activated with 5 µM ionomycin were cultured for 7 days in one of the four different culture media [Charles Rosenkrans medium (CR-1), TCM-199, potassium simplex optimization medium (KSOM) and synthetic oviductal fluid (SOF)] to study their developmental potential. The maturation rate in control, treatment 1, and treatment 2 media did not differ from each other (P > 0.05). However, the lowest degeneration of oocytes was observed in treatment 3 (P < 0.05) when compared with the other two groups. The proportion of activated oocytes was higher, while non-activated oocytes were lower in ionomycin group when compared with the group activated with ethanol (P < 0.05). The proportions of oocytes cleaved were 65.7, 56.8, 61.0 and 54.4% in CR-1, TCM-199, KSOM and SOF medium, respectively, with no significant difference. However, further development of cleaved oocytes was better in KSOM followed by SOF.

Type
Research Article
Copyright
© The Author(s), 2020. Published by Cambridge University Press

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Footnotes

*

Part of the results presented in this manuscript, were reported at the Annual Conference of the International Embryo Technology Society, 20–23 January 2018, New Orleans, USA.

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