Ultra-Structure of Bovine IVP Embryos Cultured in Two Different Serum Concentrations after Chemical Lipolysis with Forskolin.
Notice bibliographique
Résumé
Almost all media developed to support in vitro production (IVP) of bovine embryo contain fetal calf serum (FCS) and/or bovine albumin (BSA). Nevertheless, serum inhibits cleavage and development of early embryos and induces a decrease in embryo tolerance since its presence is responsible for the greater cytoplasmic lipid accumulation observed in IVP embryos. The manipulation of culture media and the use of drugs to modify embryo metabolism have been used to produce embryos with greater resistance to cryopreservation. Among these, the use of forskolin has been reported to induce an increase in pig embryo lipolysis. Therefore, the present experiment studied the ultra structure of bovine embryo produced in the presence (2.5%) or absence (0%) of FCS and forskolin. Cumulus oocyte complexes (COCs) were matured in TCM 199 (Gibco, Grand Island, NY, USA) supplemented with 10% FCS, sodium pyruvate, FSH (Pluset, Hertape Calier Saude Animal, Juatuba, Brazil), LH (Lutropin, Bioniche, Canada), penicillin and streptomycin, in a humidified atmosphere with 5% CO2 in air at 38.5°C for 24 hours. Frozen bull (Bos taurus indicus) semen from a single batch was selected through a Percoll gradient and IVF was performed in HTF (Human Tubal Fluid, Irvine Scientific, Santa Ana, California, USA) supplemented with caffeine, heparin, penicillamine, hypotaurine and in a humidified atmosphere with 5% CO2 in air at 38.5°C for 20 to 22 hours. Presumptive zygotes were cultured in SOFaa (Synthetic Oviductal Fluid) with 1% non-essential amino acids (Gibco, Grand Island, NY, USA) and 1% essential amino acids] supplemented with BSA in the presence of 2.5% FCS or in the absence of FCS. Embryos were kept in a humidified atmosphere with 5% CO2, 5% O2 and 90% N2 at 38.5°C. On day 6, embryos from each group were divided into 2 groups with or without the addition of 10 µM Forskolin. Therefore, the 4 treatments used in this study were: 2.5% FCS without forskolin, 0% FCS without forskolin, 2.5% FCS plus forskolin (2.5%+F) and 0% FCS plus forskolin (0%+F). Embryo cleavage was recorded on day three (day zero = fertilization), and blastocyst production on day 7. Transmission electron microscopy (TEM) was performed in 6 embryos from each group trough standard protocol. For statistical analysis, % cleaved and % blastocyst were transformed using the arcsine transformation (√y/100) and analyzed using ANOVA followed by Tukey test. The level of significance adopted was 5%. No differences (P>0.05) on cleavage rates were observed between groups. However a higher (P<0,05) blastocyst formation rate was obtained on groups that used FCS (2.5% FCS--46.8±13.28a and 2.5% + forskolin--46.31±11.39a) in comparison with the groups without FCS (0%FCS--21.85±8.38b and 0% + forskolin--25.71±11.58b). The ultra structural analysis showed similar features in embryos from all studied groups. However, embryos cultured in absence of FCS presented lesser and smaller lipid droplets. Moreover, embryos cultured without FCS presented more cellular debris in the perivitelinic space and in the blastocoele, indicating loss of blastomeres. The use of forskolin was not able to reduce lipid droplets in serum produced embryos. The results indicate that the removal of FCS induces a reduction in IVP embryo lipid content. In spite of this, the removal of FCS reduced embryo quality demonstrated by the increase in cellular debris and by the lower blastocyst formation rates. (poster)
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Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
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 | 0,000 | 0,000 |
| 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 | 0,001 |
| 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 découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».