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

Cryobiology. 2011 Dec;63(3):175-85. doi: 10.1016/j.cryobiol.2011.06.003. Epub 2011 Sep 3.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • 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
  • Humans
  • Mice
  • Microscopy, Electron, Scanning
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / drug effects
  • Pluripotent Stem Cells / physiology*
  • Specimen Handling
  • Sucrose / pharmacology
  • Surface Properties
  • Vitrification / drug effects

Substances

  • Cryoprotective Agents
  • Sucrose
  • Ethylene Glycol
  • Dimethyl Sulfoxide