Comparison of in vitro Maturation Media on Cattle Oocytes after in vitro Embryo Production
Sindisiwe Mbali Sithole1,2, Masindi Lottus Mphaphathi1,2, Maleke Dimpho Sebopela1,2 & Tshimangadzo Lucky Nedambale1,2 Show affiliations
- Department of Animal Sciences, Tshwane University of Technology, Faculty of Science, Private Bag X 680, Pretoria, 0001, South Africa
- Agricultural Research Council, Animal Production, Germplasm Conservation and Reproductive Biotechnologies, Private Bag X2, Irene, 0062, South Africa
American Journal of Animal and Veterinary Sciences, Volume 18, Issue 1, pp. 27–39 · doi.org/10.3844/ajavsp.2023.27.39
Abstract
The study was conducted to compare differences in vitro maturation media and their effect on subsequent in vitro embryo production. The study procedures were approved by the agricultural research council animal ethics committee (Ref no: APAEC 2020/05) and the Tshwane university of technology animal research ethics committee (Ref no: AREC 2021/08/004). Follicular fluid containing a suspension of oocytes was retrieved from ovaries through aspiration and slicing techniques. The retrieved oocytes were graded and randomly matured into four media (TCM199, Vitromat Protect, and BO IVM®) groups for 22 h, evaluated for polar body extrusion, fertilized through their respective in vitro fertilization media, and evaluated for pronucleus status. Presumptive zygotes were further cultured in vitro in their respective media, incubated for either 18 or 96 h, and evaluated for cleavage rates. The data was subjected to an appropriate analysis of variance. Student's Least Significant Differences (t-LSD) was calculated at a 5% significance level to compare means of significant treatment effects. The BO-IVM® (57.4%), TCM 199 (55.6%), and VitroMat Protect® (54.0%) recorded higher results than EGF (26.9%) for polar body formation on oocytes (P<0.05). Vitrofert® medium recorded a higher percentage (43.3%) of total fertilization rate when compared to the other media (P<0.05). VitroCleave Plus® medium recorded a higher total cleavage rate (43.3%) as compared to the other media (P<0.05). In conclusion, TCM 199, BO-IVM®, and VitroMat Protect® media rendered higher results of oocyte polar body extrusion rates compared to the EGF medium. Vitrofert® medium recorded a higher percentage (43.3%) of total fertilization rate when compared to the other media. Higher embryo development (4-8 cells and total cleavage) rates were observed in VitroCleave Plus® medium. Overall, the introduced media (VitroMat Protect®) and its media suite successfully adapted to the laboratory environment in South Africa (SA) and can therefore be adopted for optimizing the in vitro Embryo Production (IVEP) of cattle oocytes.
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Sithole, S. M., Mphaphathi, M. L., Sebopela, M. D. & Nedambale, T. L. (2023). Comparison of in vitro Maturation Media on Cattle Oocytes after in vitro Embryo Production. American Journal of Animal and Veterinary Sciences, 18(1), 27–39. https://doi.org/10.3844/ajavsp.2023.27.39
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