252 EFFECT OF FOLLICLE-STIMULATING HORMONE ADDITION ON IN VITRO MATURATION AND CLEAVAGE OF ALPACA (VICUGNA PACOS) EMBRYOS
Why this work is in the frame
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Bibliographic record
Abstract
We have previously reported that alpaca oocytes require between 38 and 42 h of maturation time (Huanca et al. 2010 Reprod. Fertil Dev. 22(1), 327). The objective of this study was to evaluate the effect of the addition of FSH in the maturation medium on nuclear maturation and cleavage rate. Ovaries were collected from a slaughterhouse and transported to the laboratory in a thermos flask containing a saline solution 0.9% with antibiotic antimycotic at 35°C. Cumulus–oocyte complexes (COC) were obtained by slicing of ovaries with a scalpel and were pooled in a conical tube for sedimentation before evaluation. The 476 COC with homogeneous cytoplasm and 2 or more layers of cumulus cells were transferred to plates with a 40-µmL drop of maturation medium TCM-199 supplemented with 10% FCS (vol:vol), 10 µg mL–1 of hCG, 0.2 mM sodium pyruvate, 50 µg mL–1 of gentamicine, and 1 µg mL–1 of oestradiol plus 0.5 µg mL–1 of FSH (Folltropin, Bioniche Animal Health, Belleville, Ontario, Canada) according the following treatments: T1 (Control): FSH by 42 h, T2: 21 h with FSH + 21 h without FSH, T3: 21 h without FSH + 21 h with FSH. The COC were maintenance under mineral oil with 10 to 12 oocytes per drop and maturated 42 h at 39°C in an atmosphere of 5% CO2 and high humidity. After the maturation time, part of the COC were removed from maturation medium and washed with PBS supplemented with 10% FCS and 1 mg/mL of hyaluronidase and fixed in ethanol: acetic acid (3:1). Oocytes were placed on the slide with minimum medium and stained with 1% orcein for 5 min. The slides were examined under a phase contrast microscope at ×400 to evaluate the status of nuclear maturation and classify as germinal vesicle (GV), metaphase I (MI), anaphase–telophase, metaphase II (MII), and degenerated. The other part of oocytes was fertilized in vitro using epididymal sperm. Motile spermatozoa were obtained by centrifugation at 600 × g on a percoll discontinuous gradient (22.5:45.0%) for 10 min. After the supernatant was removed by aspiration, the pellet was resuspended in TL HEPES and centrifuged again at 300 × g by 5 m. The pellet was resuspended in TL Stock. Gametes were coincubated for 18 h at 39°C with 5% CO2 and high humidity. Presumptive zygotes were culture in KSOM supplemented with 1 mM of glutamine, 0.3 mM of sodium pyruvate, 50 µg mL–1 of gentamicine, ethylenediaminetetraacetic acid, amino acids essentials and nonessentials, and BSA by 3 days and cultured in SOF medium for 7 days. Cleavage rate was evaluated at 72 h. Proportional data were compared by chi-square test. The proportions of oocytes reaching MII stage were 64.9 ± 8.1, 49.2 ± 9.4, and 53.8 ± 7.5% for T1, T2, and T3, respectively. Cleavage rates were 39.1, 36.3, and 33.1% and blastocysts rates were 13.6, 16.1, and 14.5% for T1, T2, and T3, respectively. The results suggest that the moment of addition of FSH does not have an effect on the maturation and cleavage rate of alpaca oocytes. This work was supported by Grant 064 – FINCyT – PIBAP 2008 and Grant 032 – PROCYT – CONCYTEC.
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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 | 0.001 | 0.001 |
| 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) | 0.000 | 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 it