The Effect of using HEPES in Culture Medium on Oocyte Maturation Rates and In vitro Fertilization in Pesisir Cattle
The Effect of using HEPES in Culture Medium on Oocyte Maturation Rates and In vitro Fertilization in Pesisir Cattle
Ferry Lismanto Syaiful*, Jaswandi Jaswandi, Mangku Mundana, Zaituni Udin
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. The 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 different 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 different 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 buffer facilitates in vitro embryo production outside the laboratory, making it a more accessible, efficient, and cost-effective option.
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