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Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR
Published on: May 29, 2014
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Gene expression analysis of human induced pluripotent stem cells cryopreserved by vitrification using StemCell Keep
Akemi Ota1, Suong-Hyu Hyon2, Shoichiro Sumi3
1BioVerde Inc., Kyoto, Japan.
Biochemistry and Biophysics Reports
|November 26, 2021
Summary
StemCell Keep (SCK) is a DMSO-free cryopreservation solution that effectively preserves human induced pluripotent stem cells (hiPS cells). SCK maintains key cell functions like anti-apoptosis and proliferation, offering a safer alternative to DMSO-based methods.
Area of Science:
- Regenerative Medicine
- Stem Cell Biology
- Cryobiology
Background:
- Advancements in regenerative medicine rely on effective cryopreservation of human somatic and induced pluripotent stem cells (hiPS cells).
- Conventional cryopreservation solutions often use dimethyl sulfoxide (DMSO), a toxic agent that can induce unwanted cell differentiation.
- Developing safer, DMSO-free alternatives is crucial for clinical applications of stem cells.
Purpose of the Study:
- To evaluate the efficacy of StemCell Keep (SCK), a DMSO-free cryoprotectant solution, for preserving hiPS cells.
- To analyze the gene expression profile of hiPS cells cryopreserved with SCK using DNA microarray analysis.
- To compare the effects of SCK with DMSO-based solutions on hiPS cell function post-thaw.
Main Methods:
- Human induced pluripotent stem cells (hiPS cells) were vitrified using either SCK or a DMSO-based solution.
- Cryopreserved hiPS cells were thawed and cultured under feeder-free conditions on Matrigel.
- RNA was extracted for DNA microarray analysis to assess gene expression.
- Gene ontology and pathway analysis (DAVID, GeneMANIA) were performed on differentially expressed genes.
Main Results:
- SCK-cryopreserved hiPS cells exhibited gene expression levels nearly equivalent to non-frozen cells.
- Key genes related to anti-apoptosis, cell adhesion, and cell proliferation were maintained at significant levels.
- Gene enrichment analysis revealed strong interactions in pathways for negative regulation of apoptosis, cell adhesion, and positive regulation of cell proliferation.
Conclusions:
- StemCell Keep (SCK) is an effective DMSO-free cryopreservation solution for hiPS cells.
- SCK successfully preserves critical hiPS cell functions, including anti-apoptosis, cell adhesion, and proliferation.
- SCK represents a promising alternative to DMSO for safe and effective stem cell cryopreservation in regenerative medicine.
