title: Slow Cooling Cryopreservation Optimized to Human Pluripotent Stem Cells.
abstract: Human pluripotent stem cells (hPSCs) have the potential for unlimited expansion and differentiation into cells that form all three germ layers. Cryopreservation is one of the key processes for successful applications of hPSCs, because it allows semi-permanent preservation of cells and their easy transportation. Most animal cell lines, including mouse embryonic stem cells, are standardly cryopreserved by slow cooling; however, hPSCs have been difficult to preserve and their cell viability has been extremely low whenever cryopreservation has been attempted.Here, we investigate the reasons for failure of slow cooling in hPSC cryopreservation. Cryopreservation involves a series of steps and is not a straightforward process. Cells may die due to various reasons during cryopreservation. Indeed, hPSCs preserved by traditional methods often suffer necrosis during the freeze-thawing stages, and the colony state of hPSCs prior to cryopreservation is a major factor contributing to cell death.It has now become possible to cryopreserve hPSCs using conventional cryopreservation methods without any specific equipment. This review summarizes the advances in this area and discusses the optimization of slow cooling cryopreservation for hPSC storage.
authors.affiliation: Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto, Japan.
authors.forename: Takamichi
authors.initials: T
authors.lastname: Miyazaki
authors.affiliation: Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan. hsuemori@frontier.kyoto-u.ac.jp.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Suemori
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D019855:Ethylene Glycol / Q000494:pharmacology; D005615:Freezing; D006801:Humans; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D013045:Species Specificity; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D047428:Protein Kinase Inhibitors; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide
keywords: Cryopreservation; Human embryonic stem cell; Human induced pluripotent stem cells; Human pluripotent stem cell; Slow-cooling; Vitrification