67 GLUTATHIONE ADDED TO CULTURE MEDIUM AND CRYOPROTECTANT SOLUTION ENHANCE THE VIABILITY OF EMBRYONIC STEM CELLS AFTER THAWING
Bibliographic record
Abstract
We have attempted to improve cell viability of embryonic stem cells (ESC) after freezing and thawing, and, as a result, an efficient cryoprotectant for cryopreserving of mouse ESC was selected in a previous study. This study was subsequently conducted to examine whether the addition of glutathione (GSH), an antioxidant, to the optimized culture medium and cryoprotectant solution could improve post-thaw cell viability of ESC. Embryonic stem cells of a mouse E14 cell line were subcultured 5 times and subsequently employed for the cryopreservation. Dulbecco’s minimal essential medium, to which 100 µm GSH was added or not, was used as a basal medium for ESC culture and a freezing solution. Dimethyl sulfoxide (10%, v/v) and ethylene glycol (10%, v/v) were used as cryoprotectants. In experiment 1, intracytoplasmic concentrations of reactive oxygen species (ROS) in ESCs subcultured and cryopreserved in GSH-free and GSH-containing medium was measured immediately after cryopreservation. A significant (P < 0.0001) decrease in ROS level was detected in ESC cultured and cryopreserved in GSH-containing medium compared with GSH-free medium. Cell viability was subsequently assessed immediately and 72 h after thawing in experiment 2. Improved post-thaw viability was detected when GSH-containing media were employed. There were no significant differences of the viability between fresh and frozen–thawed ESC, regardless of time of assessment immediately (P > 0.1325; 0.653 v. 0.604) and 72 h (P > 0.2998; 0.671 v. 0.626) after treatment. However, there was lower post-thaw viability at 0 h (P < 0.0024; 0.671 v. 0.544) and at 72 h (P < 0.0138; 0.742 v. 0.59) in ESC cultured and cryopreserved in GSH-free medium than that in ESC cultured and cryopreserved with GSH-containing medium. These results demonstrated that use of GSH as an antioxidant affects the level of ROS and promotes post-thaw survival of frozen ESC.
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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 | 0.002 | 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".