DMSO-Free Cryopreservation of Human Induced Pluripotent Stem Cells
Author Information
Author(s): Igor I. Katkov, Kan Natalia G., Cimadamore Flavio, Nelson Brandon, Snyder Evan Y., Terskikh Alexey V.
Primary Institution: CELLTRONIX, San Diego, CA, USA
Hypothesis
DMSO can be substituted with a less toxic cryoprotective agent for the cryopreservation of human pluripotent stem cells.
Conclusion
Using ethylene glycol and a programmed freezing method significantly improves the recovery of pluripotent stem cells compared to traditional methods.
Supporting Evidence
- DMSO was found to be the most toxic cryoprotectant, while ethylene glycol showed significantly lower toxicity.
- The six-step freezing protocol yielded a 63% recovery rate compared to 10-12% for standard freezing.
- Using a ROCK inhibitor during cryopreservation improved cell recovery rates.
- Cells frozen in plates showed better recovery than those frozen in clusters.
Takeaway
This study found a better way to freeze stem cells without using a harmful chemical, which helps keep them alive and healthy.
Methodology
The study compared three cryopreservation methods using different cryoprotective agents and assessed cell recovery rates after thawing.
Potential Biases
Potential bias in the assessment of cell viability due to variability in recovery measurement methods.
Limitations
The study did not explore the long-term effects of the cryopreservation methods on cell functionality.
Statistical Information
P-Value
p<0.001
Statistical Significance
p<0.001
Digital Object Identifier (DOI)
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