Rescue Potential of Supportive Embryo Culture Conditions on Bovine Embryos Derived from Metabolically Compromised Oocytes
Abstract
:1. Introduction
2. Results
2.1. The Impact of High NEFA Supplementation during IVM
2.1.1. Cumulus Cell Expansion
2.1.2. Embryo Development
2.2. Embryo Quality
2.2.1. Total Cell Count and Apoptotic Cell Index
2.2.2. Metabolic Activity of Surviving Blastocysts
2.2.3. Gene Expression Analysis of Day 8 Blastocysts
3. Discussion
3.1. High NEFA Concentrations during IVM Affect Subsequent Embryo Development but Not Quality (Apoptosis, Cellular Metabolism and Gene Expression Analysis) When Cultured in Non-Supportive Conditions (Basic SOF-BSA Medium)
3.2. Supplementation of IVC Medium with ITS Supports the Development of Lower Quality Oocytes That Otherwise Would Not Have Developed Further
3.3. Supplementation of IVC Medium with Serum Did Not Improve Developmental Competence and Quality of PA-Exposed Oocytes
3.4. Supplementation of Serum to IVC Media of HCOMBI- and BASAL-Exposed Oocytes Alters Cellular Metabolism in Surviving Blastocysts
4. Materials and Methods
4.1. Experimental Design
4.2. Preparation of NEFA Stocks and NEFA-Supplemented In Vitro Maturation Media
4.3. In Vitro Embryo Production Procedure
4.3.1. Oocyte Collection and In Vitro Maturation (IVM)
4.3.2. In Vitro Embryo Production
4.4. Outcome Parameters
4.4.1. Assessment of Cumulus Cell Expansion
4.4.2. Embryo Developmental Competence
4.4.3. Blastocyst Energy Metabolism: Determination of Pyruvate and Glucose Uptake and Lactate Production
4.4.4. Assessment of Blastocyst Cell Number and Apoptotic Cell Index
4.4.5. Blastocyst RNA Extraction, Reverse Transcription and Quantification of Gene Expression by Quantitative Polymerase Chain Reaction (qPCR)
4.5. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Maturation | Culture | Oocytes Used | Cleaved Embryos | Two-Cell Stage | Four-Cell Plus | Fragmented Embryos | Day 7 Blastocysts | Day 8 Blastocysts |
---|---|---|---|---|---|---|---|---|
BASAL | BSA | 515 | 395 a (76.69%) | 48 a§ (9.32%) | 255 a (49.51%) | 63 (12.23%) | 105 a (20.38%) | 125 a§ (24.27%) |
PA | BSA | 270 | 176 b (65.18%) | 42 b (15.55%) | 81 b (30.00%) | 35 (12.96%) | 30 b§ (11.11%) | 37 b (13.70%) |
HCOMBI | BSA | 527 | 377 ab (71.53%) | 63 ab§ (11.95%) | 221 c (41.93%) | 65 (12.33%) | 87 ab§ (16.50%) | 96 ab§ (18.21%) |
BASAL | ITS | 510 | 406 §$ (79.6%) | 63 (12.35%) | 250 x (49.01%) | 56§ (10.98%) | 124 (24.31%) | 139 (27.25%) |
PA | ITS | 278 | 200 $ (71.94%) | 43 (15.46%) | 110 y (39.56%) | 30 (10.79%) | 52 (18.7%) | 68 (24.46%) |
HCOMBI | ITS | 525 | 387 § (73.71%) | 46 (8.76%) | 235 xy (44.76%) | 84 § (16.00%) | 101 (19.23%) | 120 (22.85%) |
BASAL | Serum | 488 | 348 jk§ (71.31%) | 73 j (14.95%) | 182 j (37.29%) | 55 (11.27%) | 119 j (24.39%) | 136 j (27.86%) |
PA | Serum | 247 | 156 j§ (63.15%) | 54 k (21.86%) | 53 k (21.45%) | 30 (12.14%) | 30 k (12.14%) | 45 k (18.21%) |
HCOMBI | Serum | 377 | 275 k (72.94%) | 55 jk (14.58%) | 149 j (39.52%) | 41 (10.87%) | 88 j (23.34%) | 91 jk (24.13%) |
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Smits, A.; Leroy, J.L.M.R.; Bols, P.E.J.; De Bie, J.; Marei, W.F.A. Rescue Potential of Supportive Embryo Culture Conditions on Bovine Embryos Derived from Metabolically Compromised Oocytes. Int. J. Mol. Sci. 2020, 21, 8206. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21218206
Smits A, Leroy JLMR, Bols PEJ, De Bie J, Marei WFA. Rescue Potential of Supportive Embryo Culture Conditions on Bovine Embryos Derived from Metabolically Compromised Oocytes. International Journal of Molecular Sciences. 2020; 21(21):8206. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21218206
Chicago/Turabian StyleSmits, Anouk, Jo L. M. R. Leroy, Peter E. J. Bols, Jessie De Bie, and Waleed F. A. Marei. 2020. "Rescue Potential of Supportive Embryo Culture Conditions on Bovine Embryos Derived from Metabolically Compromised Oocytes" International Journal of Molecular Sciences 21, no. 21: 8206. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21218206