Study of the Metabolomics of Equine Preovulatory Follicular Fluid: A Way to Improve Current In Vitro Maturation Media
Production of equine embryos in vitro is currently a commercial technique and a reliable way of obtaining offspring. In order to produce those embryos, immature oocytes are retrieved from postmortem ovaries or live mares by ovum pick-up (OPU), matured in vitro (IVM), fertilized by intracytoplasmic s...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-05-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/10/5/883 |
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| author | Pablo Fernández-Hernández María Jesús Sánchez-Calabuig Luis Jesús García-Marín María J. Bragado Alfonso Gutiérrez-Adán Óscar Millet Chiara Bruzzone Lauro González-Fernández Beatriz Macías-García |
| author_facet | Pablo Fernández-Hernández María Jesús Sánchez-Calabuig Luis Jesús García-Marín María J. Bragado Alfonso Gutiérrez-Adán Óscar Millet Chiara Bruzzone Lauro González-Fernández Beatriz Macías-García |
| author_sort | Pablo Fernández-Hernández |
| collection | DOAJ |
| description | Production of equine embryos in vitro is currently a commercial technique and a reliable way of obtaining offspring. In order to produce those embryos, immature oocytes are retrieved from postmortem ovaries or live mares by ovum pick-up (OPU), matured in vitro (IVM), fertilized by intracytoplasmic sperm injection (ICSI), and cultured until day 8–10 of development. However, at best, roughly 10% of the oocytes matured in vitro and followed by ICSI end up in successful pregnancy and foaling, and this could be due to suboptimal IVM conditions. Hence, in the present work, we aimed to elucidate the major metabolites present in equine preovulatory follicular fluid (FF) obtained from postmortem mares using proton nuclear magnetic resonance spectroscopy (<sup>1</sup>H-NMR). The results were contrasted against the composition of the most commonly used media for equine oocyte IVM: tissue culture medium 199 (TCM-199) and Dulbecco’s modified eagle medium/nutrient mixture F-12 Ham (DMEM/F-12). Twenty-two metabolites were identified in equine FF; among these, nine of them are not included in the composition of DMEM/F-12 or TCM-199 media, including (mean ± SEM): acetylcarnitine (0.37 ± 0.2 mM), carnitine (0.09 ± 0.01 mM), citrate (0.4 ± 0.04 mM), creatine (0.36 ± 0.14 mM), creatine phosphate (0.36 ± 0.05 mM), fumarate (0.05 ± 0.007 mM), glucose-1-phosphate (6.9 ± 0.4 mM), histamine (0.25 ± 0.01 mM), or lactate (27.3 ± 2.2 mM). Besides, the mean concentration of core metabolites such as glucose varied (4.3 mM in FF vs. 5.55 mM in TCM-199 vs. 17.5 mM in DMEM/F-12). Hence, our data suggest that the currently used media for equine oocyte IVM can be further improved. |
| first_indexed | 2024-03-10T19:43:34Z |
| format | Article |
| id | doaj.art-2d29c0c548e9497eb78c45bb7fc59b01 |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-10T19:43:34Z |
| publishDate | 2020-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-2d29c0c548e9497eb78c45bb7fc59b012023-11-20T00:58:48ZengMDPI AGAnimals2076-26152020-05-0110588310.3390/ani10050883Study of the Metabolomics of Equine Preovulatory Follicular Fluid: A Way to Improve Current In Vitro Maturation MediaPablo Fernández-Hernández0María Jesús Sánchez-Calabuig1Luis Jesús García-Marín2María J. Bragado3Alfonso Gutiérrez-Adán4Óscar Millet5Chiara Bruzzone6Lauro González-Fernández7Beatriz Macías-García8Research Group of Intracellular Signaling and Technology of Reproduction, Research Institute INBIO G + C, University of Extremadura, 10003 Cáceres, SpainDepartment of Animal Reproduction, INIA, 28040 Madrid, SpainResearch Group of Intracellular Signaling and Technology of Reproduction, Research Institute INBIO G + C, University of Extremadura, 10003 Cáceres, SpainResearch Group of Intracellular Signaling and Technology of Reproduction, Research Institute INBIO G + C, University of Extremadura, 10003 Cáceres, SpainDepartment of Animal Reproduction, INIA, 28040 Madrid, SpainPrecision Medicine and Metabolism Lab., CICbioGUNE, 48160 Vizcaya, SpainPrecision Medicine and Metabolism Lab., CICbioGUNE, 48160 Vizcaya, SpainResearch Group of Intracellular Signaling and Technology of Reproduction, Research Institute INBIO G + C, University of Extremadura, 10003 Cáceres, SpainResearch Group of Intracellular Signaling and Technology of Reproduction, Research Institute INBIO G + C, University of Extremadura, 10003 Cáceres, SpainProduction of equine embryos in vitro is currently a commercial technique and a reliable way of obtaining offspring. In order to produce those embryos, immature oocytes are retrieved from postmortem ovaries or live mares by ovum pick-up (OPU), matured in vitro (IVM), fertilized by intracytoplasmic sperm injection (ICSI), and cultured until day 8–10 of development. However, at best, roughly 10% of the oocytes matured in vitro and followed by ICSI end up in successful pregnancy and foaling, and this could be due to suboptimal IVM conditions. Hence, in the present work, we aimed to elucidate the major metabolites present in equine preovulatory follicular fluid (FF) obtained from postmortem mares using proton nuclear magnetic resonance spectroscopy (<sup>1</sup>H-NMR). The results were contrasted against the composition of the most commonly used media for equine oocyte IVM: tissue culture medium 199 (TCM-199) and Dulbecco’s modified eagle medium/nutrient mixture F-12 Ham (DMEM/F-12). Twenty-two metabolites were identified in equine FF; among these, nine of them are not included in the composition of DMEM/F-12 or TCM-199 media, including (mean ± SEM): acetylcarnitine (0.37 ± 0.2 mM), carnitine (0.09 ± 0.01 mM), citrate (0.4 ± 0.04 mM), creatine (0.36 ± 0.14 mM), creatine phosphate (0.36 ± 0.05 mM), fumarate (0.05 ± 0.007 mM), glucose-1-phosphate (6.9 ± 0.4 mM), histamine (0.25 ± 0.01 mM), or lactate (27.3 ± 2.2 mM). Besides, the mean concentration of core metabolites such as glucose varied (4.3 mM in FF vs. 5.55 mM in TCM-199 vs. 17.5 mM in DMEM/F-12). Hence, our data suggest that the currently used media for equine oocyte IVM can be further improved.https://www.mdpi.com/2076-2615/10/5/883IVMoocytesequinemetabolomic |
| spellingShingle | Pablo Fernández-Hernández María Jesús Sánchez-Calabuig Luis Jesús García-Marín María J. Bragado Alfonso Gutiérrez-Adán Óscar Millet Chiara Bruzzone Lauro González-Fernández Beatriz Macías-García Study of the Metabolomics of Equine Preovulatory Follicular Fluid: A Way to Improve Current In Vitro Maturation Media Animals IVM oocytes equine metabolomic |
| title | Study of the Metabolomics of Equine Preovulatory Follicular Fluid: A Way to Improve Current In Vitro Maturation Media |
| title_full | Study of the Metabolomics of Equine Preovulatory Follicular Fluid: A Way to Improve Current In Vitro Maturation Media |
| title_fullStr | Study of the Metabolomics of Equine Preovulatory Follicular Fluid: A Way to Improve Current In Vitro Maturation Media |
| title_full_unstemmed | Study of the Metabolomics of Equine Preovulatory Follicular Fluid: A Way to Improve Current In Vitro Maturation Media |
| title_short | Study of the Metabolomics of Equine Preovulatory Follicular Fluid: A Way to Improve Current In Vitro Maturation Media |
| title_sort | study of the metabolomics of equine preovulatory follicular fluid a way to improve current in vitro maturation media |
| topic | IVM oocytes equine metabolomic |
| url | https://www.mdpi.com/2076-2615/10/5/883 |
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