49 IN VITRO DEVELOPMENT OF PORCINE CLONED EMBRYOS FOLLOWING DIFFERENT ACTIVATION TREATMENTS
Bibliographic record
Abstract
The present study compared the development of cloned porcine embryos following different activation treatments. Cumulus-oocyte complexes (COCs) were aspirated from slaughterhouse ovaries and cultured for 22 h in NCSU#23 medium supplemented with 10% porcine follicular fluid, 0.57 mM cysteine, 0.5 µg mL-1 LH, 0.5 µg mL-1 FSH and 10 ng mL-1 EGF. The COCs were further cultured for an additional 22 h in the same medium at 39°C in an atmosphere of 5% CO2 in air, without hormonal supplements. Primary cultures of fibroblasts from a female fetus on Day 40 of gestation were established in DMEM + 15% FCS. For nuclear donation, cells at the 5th-6th passage were cultured in DMEM + 0.5% FCS for 5 days in order to arrest the cells in G0/G1. Following enucleation, oocytes were reconstructed by transfer of donor cells and fusion by means of three DC pulses (1.4 kV cm-1, 30 µs) delivered by a BTX 200, in 0.28 M mannitol containing 0.01 mM CaCl2 and 0.01 mM MgCl2. Eggs were then divided into three treatment groups; control (without further treatment, Group 1), eggs cultured in 10 µg mL-1 cycloheximide (CHX) for 5 h (Group 2), and eggs cultured in 1.9 mM 6-dimethylaminopurine (6-DMAP) for 5 h (Group 3). The eggs were then cultured in sets of 30 in 60 µL drops of NCSU#23 supplemented with 4 µg mL BSA (essentially fatty acid free) until Day 7 at 39°C in a humidified atmosphere of 5% CO2. On Day 4 the culture were fed by adding 20 µL NCSU#23 supplemented with 10% FBS. All experiments were performed as 4 replicates and statistical analysis was performed by one-way ANOVA (P < 0.05). Blastocyst development rates were significantly higher (P < 0.05) in Group 3 embryos compared to Group 1 controls (27.6 ± 2.7% v. 20.1 ± 4.1%, respectively), but rates did not differ in Group 2 (23.8 ± 5.7%) compared to control. Total cell number in Group 3 blastocysts was, however, significantly higher (P < 0.05) than in Groups 1 and 2 (44.6 ± 2.4 v. 19.9 ± 1.9 and 21.9 ± 2.1, respectively). These results suggest that 6-DMAP is more efficient than cycloheximide in the activation of electrically fused NT oocytes during in vitro production of cloned porcine embryos. [Supported by High Technology Development Project for Agriculture and Forestry Korea, MAF-SGRP, 300012-05-3-SB010 and Cho-A Pharm. Co. LTD.]
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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 | 0.000 |
| 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".