Slow freezing cryopreservation of Korean bovine blastocysts with an additional sucrose pre-equilibration step.
| Autor: | Jung S; Veterinary Medical Center and College of Veterinary Medicine, Laboratory of Veterinary Embryology and Biotechnology (VETEMBIO), Chungbuk National University, Cheongju, Republic of Korea.; ET Biotech Co. Ltd., Jangsu, Republic of Korea., Sul H; ET Biotech Co. Ltd., Jangsu, Republic of Korea., Oh D; Veterinary Medical Center and College of Veterinary Medicine, Laboratory of Veterinary Embryology and Biotechnology (VETEMBIO), Chungbuk National University, Cheongju, Republic of Korea.; Institute of Stem Cell and Regenerative Medicine (ISCRM), Chungbuk National University, Cheongju, Republic of Korea., Jung YG; ET Biotech Co. Ltd., Jangsu, Republic of Korea., Lee J; Department of Companion Animal Industry, Semyung University, Jecheon, Republic of Korea., Hyun SH; Veterinary Medical Center and College of Veterinary Medicine, Laboratory of Veterinary Embryology and Biotechnology (VETEMBIO), Chungbuk National University, Cheongju, Republic of Korea.; Institute of Stem Cell and Regenerative Medicine (ISCRM), Chungbuk National University, Cheongju, Republic of Korea.; Graduate School of Veterinary Biosecurity and Protection, Chungbuk National University, Cheongju, Republic of Korea. |
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| Jazyk: | angličtina |
| Zdroj: | Frontiers in veterinary science [Front Vet Sci] 2024 Apr 10; Vol. 11, pp. 1400899. Date of Electronic Publication: 2024 Apr 10 (Print Publication: 2024). |
| DOI: | 10.3389/fvets.2024.1400899 |
| Abstrakt: | Introduction: Embryo cryopreservation is a valuable technique used for preserving genetic resources for long periods. However, the survival rate of embryos is dependent on the method used. Therefore, in this study, we evaluated the efficiency of slow freezing method but with an additional dehydration step prior to freezing to overcome the formation of ice crystals. Methods: Oocytes collected from the ovaries of native Korean cattle subjected to in vitro fertilization were cultured for 7 days until the formation of expanded blastocysts. Before freezing, the blastocysts were placed in four pre-equilibration media: a control medium with no addition of sucrose, and three experimental media with the addition of 0.1, 0.25, and 0.5 M sucrose, respectively. Then, the pre-equilibrated embryos were frozen. Embryo survival and hatching rates were evaluated morphologically at 24, 48, and 72 h after thawing. Immunofluorescence staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, and gene expression analysis of the re-expanded blastocytes were examined 24 h after freeze-thawing. Results: The survival rate was significantly higher in the 0.1 M group than in the control group ( p < 0.05), and the hatching rate at 72 h was significantly higher in the 0.25 and 0.5 M groups than in the control group ( p < 0.05). TUNEL-positive cells were significantly lower in the 0.25 M group than in the control group (12.5 ± 0.9 vs. 8.3 ± 0.8; p < 0.05). The gene expression of BCL2 associated X, heat shock protein 70 kDa, and aquaporin 3 in the 0.25 M group was significantly lower than that in the control group ( p < 0.05). Conclusion: Our study revealed that treatment with 0.25 M sucrose before slow freezing improved the viability of bovine embryos after freeze-thawing. Competing Interests: SJ, HS, and Y-GJ were employed by ET Biotech Co. Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2024 Jung, Sul, Oh, Jung, Lee and Hyun.) |
| Databáze: | MEDLINE |
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