138 THE EFFECT OF SYNTHETIC HYALURONAN, BSA AND SERUM ON IN VITRO DEVELOPMENT AND GENE TRANSCRIPTION OF BOVINE BLASTOCYSTS
Bibliographic record
Abstract
The objective of this study was to examine the effect of synthetic hyaluronan (s-HA), BSA and fetal calf serum (FCS) on bovine embryo in vitro development, ultrastructure, and mRNA transcription of four developmentally important genes: apoptosis (BAX), oxidative stress (SOX), growth factor (IGF-II), and cell-to-cell adhesion (Ecad). A total of 1406 presumptive zygotes (7 replicates) were cultured initially in two Groups: 1, SOFaa + 4% BSA only, and 2, SOFaa + 4% BSA and 10% FCS. On Day 4 (96 h after insemination) of culture, the number of zygotes that developed to the <8-cell-stage were recorded, and 2.5 mg/mL of s-HA (MAP-5; Bioniche Inc, Belleville, ON, Canada) was added to half of the embryos from each group; the other half received an equivalent volume of corresponding SOF medium without s-HA. Embryos were cultured in 50-µL drops (25 zygotes per drop) under paraffin oil at 39°C and 5% CO2 in humidified air. Cleavage rates were recorded on Day 2 and the number of blastocysts on Days 7, 8, and 9. At least five blastocysts from each replicate and from each treatment were frozen for evaluation of gene expression patterns. Poly(A) mRNA was prepared from 4–5 groups of pools of 10 embryos. The quantification of all gene transcripts was performed by real time quantitative RT-PCR in three replicates. The fine structure of blastocysts was studied using transmission electron microscopy. Embryo developmental stages and blastocyst formation were analyzed by chi-square analysis, and data on mRNA expression were analyzed by one-way repeated-measures ANOVA. No differences in cleavage rates were observed between groups. There was no difference between the BSA group with or without s-HA in the percentages of embryos developed to the blastocyst stage at Days 7, 8, and 9 (38.3 and 38.1%, respectively). However, significantly (P < 0.05) less blastocysts developed in medium supplemented with BSA + FCS (18.3%) or with BSA + FCS + s-HA (27.4%). Synthetic HA added to the medium containing BSA significantly (P < 0.05) increased the level of expression of EGF-II and decreased (P < 0.05) the level of expression of BAX, SOX, and Ecad. On the other hand, presence of FCS significantly (P < 0.05) increased the level of SOX and decreased the level of IGF-II (P < 0.05), and the addition of s-HA to SOF containing FCS showed no effect on the level of transcription of any analyzed genes. The general fine structure of embryos cultured with s-HA regardless of protein supplement was conspicuously improved in comparison with the respective controls. It can be concluded that, within our culture system, addition of s-HA on Day 4 of culture to the SOFaa medium supplemented with BSA but not in combination with FCS showed a positive effect on embryo development and molecular composition of the embryos.
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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 | 0.001 | 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) | 0.000 | 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".