213 <i>IN VITRO</i> DEVELOPMENT OF BOVINE EMBRYOS IN THE PRESENCE OF GROWTH HORMONE, INSULIN-LIKE GROWTH FACTOR-1, AND INSULIN IN OOCYTE MATURATION AND EMBRYO CULTURE MEDIA
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Notice bibliographique
Résumé
The addition of hormones and growth factors to bovine IVM and IVC media has been reported to affect early embryonic development by enhancing the blastocyst formation rate and quality of embryos produced. The purpose of this study was to investigate the influence of adding growth hormone (GH), insulin-like growth factor-1 (IGF-1), and insulin to IVM and IVC media. Blastocyst production rate and blastocyst quality, as verified by the number of cells with DNA fragmentation, were evaluated. Ovaries from an abattoir were transported to the laboratory and COC were selected and cultured in IVM medium 199 (Earle's salts, Sigma, St. Louis, MO, USA), 10% fetal calf serum (Sigma), 50 µg mL–1 of sodium pyruvate, 1 µg mL–1 of estradiol (Sigma), 50 µg mL–1 of hCG (Profasi hp®, 5000 IU, Serono Inc., Rockland, MA, USA), 5 µg mL–1 of FSH (Folltropin®, Vetrepharm, Ontario, Canada), and 75 µg mL–1 of gentamicin sulfate for 24 h. After IVF (18 h), zygotes were partially denuded and transferred to IVC medium HTF (HTF®, Irvine Scientific, Santa Ana, CA, USA) and BME (BME®, Sigma), in a 1:1 proportion (HTF:BME), 0.6% BSA (Sigma), 0.01% myoinositol (Sigma), and 75 µg mL–1 of gentamicin sulfate, at 38.5°C, in a humidified atmosphere of 5% CO2 in air, supplemented with 10% fetal calf serum at Day 3 of culture. Three different experiments were performed. The first and second experiments were analyzed using the chi-square test ( P &lt; 0.05). The third experiment was analyzed with the general linear model of SAS® (SAS Institute Inc., Cary, NC, USA) and the Tukey test ( P &lt; 0.1). In the first experiment, oocytes were cultured in IVM medium supplemented with GH (10 ng mL–1), IGF-1 (100 ng mL–1), insulin (1 µg mL–1), or all 3 combined. In the second experiment, IVC medium was supplemented with GH, IGF-1, insulin, or all 3 combined (same concentrations as above). In the third experiment, the quality of the embryos produced in the first 2 experiments was determined by the percentage of cells with DNA fragmentation. After 96 h of culture, embryos were stained with orange acridin (100 µg mL–1) and propidium iodide (100 µg mL–1) and slides were evaluated by fluorescence microscopy (450 to 490 nm). Rates of blastocyst production (blastocysts/oocytes) in the first experiment (29, 28, 28, 26, and 28% for control, GH, IGF-1, insulin, or all 3 combined, respectively) and in the second experiment (35, 35, 36, 35, and 31%) were not statistically different among the groups. In the third experiment, the addition of GH, IGF-1, or insulin to IVM medium did not affect the DNA fragmentation rate (11, 5, 2, 12, and 12%). However, the addition of insulin to IVC medium led to a higher DNA fragmentation rate (24%), when compared with the other groups (11, 10, 6, and 8% for control, GH, IGF-1, and all 3 combined). The addition of GH or IGF-1 to bovine IVM and IVC media did not affect the blastocyst production rate or the quality of embryos produced. The quality of embryos cultured in the presence of insulin was negatively affected.
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Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,003 | 0,001 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découle