Improvement on the viability of preantral Bos indicus follicles cultured in vitro by addition of follicle stimulating hormone (FSH)
Bibliographic record
Abstract
The aim of this study was to evaluate the effect of FSH addition in the medium for in vitro culture of preantral follicles of Bos indicus females. Ovaries (n=10) were collected at a local slaughterhouse from five adult cycling Bos indicus cows (Nelore) with body condition score from 2.5 to 3.5 (range 0-5). After collection, ovaries were washed in 70% ethanol and PBS (Embriolife®, Vitrocell, Brazil). The surrounding tissue was removed and ovaries were sectioned longitudinally. The ovarian cortex was divided in 3x3x1 mm fragments. One fragment per animal was immediately fixed in Bouin (non-cultured control, D0). The other fragments (n=8) were individually cultured in 24- well culture plates containing 1 ml of minimum essential medium (MEM, Gibco BRL, Rockville, MD, USA; osmolarity 300 mOsm/l, pH 7.2) supplemented (MEM+) with ITS (6.25 mg/ml insulin, 6.25 mg/ml transferrin, and 6.25 ng/ml selenium), 0.23 mM pyruvate, 2 mM glutamine, 2 mM hypoxantina, 1.25 mg/ml bovine serum albumin (BSA Gibco BRL, Rockville, MD, USA), 20 IU/ml penicillin and 200 mg/ml streptomycin. Unspecified reagents were obtained from Sigma (St. Louis, MO, USA). MEM + medium was added to different concentrations (50, 100 and 200 ng/ml) FSH (Folltropin®, Bioniche Canada Inc, Ontario, Canada). The in vitro culture medium was tested during two (D2) or six (D6) days. Every two days, the culture media was replaced by fresh aliquots. For the analysis of integrity and degree of follicular development we used histology with periodic acid-Schiff (PAS) and hematoxylin staining. The classification was based on the evaluation of follicular development stage (primordial, primary and secondary) and morphological integrity as normal or degenerated. Data were submitted to ANOVA (p ≤ 0.05). We evaluated 2,250 preantral follicles (normal or degenerated), being 772 primordial and 1,478 in development. After two days of culture, FSH at 100 ng/ml provided a higher proportion (51.2%; 128/250) of follicles morphologically intact when compared to the other groups: 27.2% (68/250) to MEM; 30.4% (76/250) to 50 ng/ml; 45.2% (113/250) to 200 ng/ml (p 0.05). We conclude that MEM+ supplemented with 100 ng/ml FSH for two or six days of in vitro culture was the most effective treatment to provide development and morphological integrity of preantral follicles from Bos indicus cows.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| 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 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.001 | 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 source (direct Gemma or distilled Codex), 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".