Ultra-Structure of Bovine IVP Embryos Cultured in Two Different Serum Concentrations after Chemical Lipolysis with Forskolin.
Bibliographic record
Abstract
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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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.001 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".