title: Rationally optimized cryopreservation of multiple mouse embryonic stem cell lines: I--Comparative fundamental cryobiology of multiple mouse embryonic stem cell lines and the implications for embryonic stem cell cryopreservation protocols.
abstract: The post-thaw recovery of mouse embryonic stem cells (mESCs) is often assumed to be adequate with current methods. However as this publication will show, this recovery of viable cells actually varies significantly by genetic background. Therefore there is a need to improve the efficiency and reduce the variability of current mESC cryopreservation methods. To address this need, we employed the principles of fundamental cryobiology to improve the cryopreservation protocol of four mESC lines from different genetic backgrounds (BALB/c, CBA, FVB, and 129R1 mESCs) through a comparative study characterizing the membrane permeability characteristics and membrane integrity osmotic tolerance limits of each cell line. In the companion paper, these values were used to predict optimal cryoprotectants, cooling rates, warming rates, and plunge temperatures, and then these predicted optimal protocols were validated against standard freezing protocols.
authors.affiliation: Department of Biological Sciences, Northern Illinois University, DeKalb, IL 60115, USA. Electronic address: ckashuba@niu.edu.
authors.forename: Corinna M
authors.initials: CM
authors.lastname: Kashuba
authors.affiliation: Department of Mathematical Sciences, Northern Illinois University, DeKalb, IL 60115, USA.
authors.forename: James D
authors.initials: JD
authors.lastname: Benson
authors.affiliation: University of Missouri Comparative Medicine Center, 4011 Discovery Drive, Columbia, MO 65201, USA.
authors.forename: John K
authors.initials: JK
authors.lastname: Critser
mesh_terms: D000818:Animals; D002460:Cell Line; D002463:Cell Membrane Permeability; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D053595:Embryonic Stem Cells; D005615:Freezing; D051379:Mice; D009995:Osmosis
chemical_list: D003451:Cryoprotective Agents
keywords: Cryopreservation; ES cells; Embryonic stem; Embryonic stem cells; Fundamental cryobiology; Membrane permeability; Mouse; Osmotic tolerance limits; Osmotically inactive cell volume