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The efect of using HEPES in Culture Medium on Oocyte Maturation Rates and In vitro Fertilization in Pesisir Cattle
Author(s):
1. Ferry Lismanto Syaiful: Department of Animal Production Technology, Faculty of Animal Science, Universitas Andalas, Padang-West Sumatra, Indonesia.
2. Jaswandi Jaswandi: Department of Animal Production Technology, Faculty of Animal Science, Universitas Andalas, Padang-West Sumatra, Indonesia
3. Mangku Mundana: Department of Animal Production Technology, Faculty of Animal Science, Universitas Andalas, Padang-West Sumatra, Indonesia
4. Zaituni Udin: Department of Animal Production Technology, Faculty of Animal Science, Universitas Andalas, Padang-West Sumatra, Indonesia
Abstract:
This research aims to assess oocyte maturation rates, in vitro fertilization, and pronuclei development in Pesisir cattle, and to determine the optimal culture medium and culture duration for in vitro fertilization techniques. hTe culture media used include TCM-199 medium supplemented with both B-O (Brackett Oliphant) medium (HEPES, without the use of 5% CO2) and modified B-O (mB-O) medium (without HEPES, with 5% CO2). For the oocyte maturation stage, the culture period was 24 hours, while for in vitro fertilization, various culture periods (6, 12, 18h) were utilized. The analysis of research data acquisition was conducted using a Completely Randomized Design, utilizing SPSS. The results of in vitro maturation of Pesisir cattle oocytes indicate the development of GV (Germinal Vesicle), GVBD (Germinal Vesicle Breakdown), M-I (Maturation-I) and M-II (Maturation-II) cells in diferent culture media as follows: B-O medium (14.29%, 0%, 33.33%, 57.14%), mB-O (17.65%, 0%, 23.53%, 58.82%). The results of in vitro fertilization rates using diferent culture media at various incubation periods were as follows: B-O medium at 6, 12, and 18h yields 57.14%, 52.17%, and 48.65%, while mB-O medium yields 57.69%, 51.22%, and 48.28%. The research concludes that the addition of HEPES in the B-O medium can substitute for the role of 5% CO2 in the mB-O medium and even enhance oocyte maturation and in vitro fertilization in Pesisir cattle. This demonstrates that using HEPES bufer facilitates in vitro embryo production outside the laboratory, making it a more accessible, eficient, and cost-efective option.
Page(s): 450-457
Published: Journal: Journal of animal health and production, Volume: 12, Issue: 3, Year: 2024
Keywords:
in vitro fertilization , Oocyte maturation , Brackett Oliphant , 5 CO2 , Pronucleus , HEPES
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