106 Efficiency of an in-house developed medium for bovine in vitro embryo production

The commercial use of ovum pickup and in vitro embryo production (OPU-IVEP) in bovines has increased worldwide, improving reproductive performance and genetic gain. However, the acceptance of the technology in many dairy-producing developing nations is nascent owing to the high cost, which is mainly due to the imported media. The present study assessed the efficacy of an in-house developed cost-effective media suite (comprising IVM, IVF, IVC, and Wash media) with specific additives for bovine IVEP compared with commercially available bovine IVEP media. The in-house medium was developed at Indian Immunologicals Ltd., Hyderabad. The experiments were performed simultaneously in two setups (i.e. at SLU, Sweden), using slaughterhouse ovaries as the source of oocytes, and at NDDB, India, using OPU. At SLU, 157 and 167 oocytes were subjected to IVEP using in-house and control media suites, respectively. Further, the embryos developed on Day 8 from IVF in both groups were subjected to nuclei count analysis using confocal microscopy with DAPI staining. At NDDB, 80 and 91 OPUs were performed using the control and in-house media suites, respectively. IVEP using commercial media was performed as per the manufacturer’s protocol. In the test media, oocytes were subjected to IVM (22 h in 5% CO₂ in air at 38.5°C with maximum humidity), IVF (22 h in 5% CO₂ in air at 38.5°C with maximum humidity), and IVC (7 days from IVF in 5% CO₂, 5% O₂, and 90% N₂ at maximum humidity at 38.5°C). The cleavage and blastocyst rates were calculated by dividing the number of oocytes cleaved and blastocysts developed by the total number of oocytes in IVC, respectively. Descriptive statistics were calculated for cleavage rate and blastocyst rate for both setups, embryos/OPU at NDDB, and nuclei count at SLU for control and test media, and means are represented as mean ± s.e.m. Welch two-sample t-tests were used for comparing different means between the media using RStudio (version 2024.04.1+748). At SLU, the blastocyst rates were 25.9 ± 2.6% and 28.1 ± 4.7% in test and control media, respectively. Blastocyst rates did not differ significantly between the test and control media. The mean nuclei count was significantly higher in the control group (test: 67.4 vs. control: 89.2; P = 0.004), suggesting slightly enhanced cell proliferation in the control media. However, the high standard deviation in the control group (test: 24.3 vs. control: 42.9) indicates the stability of the test media. At NDDB, no significant difference was found between test and control media in terms of cleavage rate (66.2 ± 2.7% vs. 65.0 ± 2.7%; P = 0.188), blastocyst rate (20.6 ± 2.1% vs. 24.6 ± 2.3%; P = 0.925), or embryos/OPU (3.4 ± 0.4 vs. 4.6 ± 0.6; P = 0.66). The study reveals the efficacy of in-house developed media suites in both slaughterhouse and OPU-IVEP systems, with comparable results indicating the suitability of the media to be used in bovine IVEP.