title: Effective surface-based cryopreservation of human embryonic stem cells by vitrification.
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.
authors.affiliation: Fraunhofer Institute of Biomedical Engineering, St. Ingbert, Germany.
authors.forename: A F J
authors.initials: AF
authors.lastname: Beier
authors.forename: J C
authors.initials: JC
authors.lastname: Schulz
authors.forename: D
authors.initials: D
authors.lastname: Dörr
authors.forename: A
authors.initials: A
authors.lastname: Katsen-Globa
authors.forename: A
authors.initials: A
authors.lastname: Sachinidis
authors.forename: J
authors.initials: J
authors.lastname: Hescheler
authors.forename: H
authors.initials: H
authors.lastname: Zimmermann
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D019855:Ethylene Glycol / Q000494:pharmacology; D061252:Feeder Cells / Q000502:physiology; D005434:Flow Cytometry; D005615:Freezing; D006801:Humans; D051379:Mice; D008855:Microscopy, Electron, Scanning; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D013048:Specimen Handling; D013395:Sucrose / Q000494:pharmacology; D013499:Surface Properties; D058989:Vitrification / Q000187:drug effects
chemical_list: D003451:Cryoprotective Agents; D013395:Sucrose; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide