Effective surface-based cryopreservation of human embryonic stem cells by vitrification

► We established a surface-based protocol for bulk vitrification of hESCs. ► It results in very high recovery rates and maintains pluripotency. ► Method is easy to handle and allows exact incubation times. ► Slow rate protocol of adherent hESCs results in disturbed membrane integrity. ► Efficiency o...

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Bibliographic Details
Published in:Cryobiology 2011-12, Vol.63 (3), p.175-185
Main Authors: Beier, A.F.J., Schulz, J.C., Dörr, D., Katsen-Globa, A., Sachinidis, A., Hescheler, J., Zimmermann, H.
Format: Article
Language:English
Subjects:
Adherent cryopreservation
Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy
Animals
Applied cell therapy and gene therapy
Biological and medical sciences
Cell Culture Techniques
Cell Differentiation - drug effects
Cell Differentiation - physiology
Cell Line
Cell Survival - drug effects
Cell Survival - physiology
Cryopreservation - methods
Cryoprotective Agents - pharmacology
Dimethyl Sulfoxide - pharmacology
Embryonic Stem Cells - cytology
Embryonic Stem Cells - drug effects
Embryonic Stem Cells - physiology
Ethylene Glycol - pharmacology
Feeder Cells - physiology
Flow Cytometry
Freezing
Human embryonic stem cells
Humans
Medical sciences
Mice
Microscopy, Electron, Scanning
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - drug effects
Pluripotent Stem Cells - physiology
Specimen Handling
Sucrose - pharmacology
Surface Properties
Transfusions. Complications. Transfusion reactions. Cell and gene therapy
Vitrification
Vitrification - drug effects
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Summary:► We established a surface-based protocol for bulk vitrification of hESCs. ► It results in very high recovery rates and maintains pluripotency. ► Method is easy to handle and allows exact incubation times. ► Slow rate protocol of adherent hESCs results in disturbed membrane integrity. ► Efficiency of slow rate protocol is significantly lower and depends on colony size. Human embryonic stem cells (hESCs) are candidates for many applications in the areas of regenerative medicine, tissue engineering, basic scientific research as well as pharmacology and toxicology. However, use of hESCs is limited by their sensitivity to freezing and thawing procedures. Hence, this emerging science needs new, reliable preservation methods for the long-term storage of large quantities of functional hESCs remaining pluripotent after post-thawing and culturing. Here, we present a highly efficient, surface based vitrification method for the cryopreservation of large numbers of adherent hESC colonies, using modified cell culture substrates. This technique results in much better post-thaw survival rate compared to cryopreservation in suspension and allows a quick and precise handling and storage of the cells, indicating low differentiation rates.
ISSN:0011-2240
1090-2392
DOI:10.1016/j.cryobiol.2011.06.003