title: The ROCK inhibitor Y-27632 enhances the survival rate of human embryonic stem cells following cryopreservation.
abstract: After slow freezing, the survival rate of human embryonic stem (hES) cells is poor and inconsistent. The aim of this study was to increase the freeze-thaw survival rate of hES cells by utilizing the ROCK inhibitor Y-27632. hES cell colonies were first treated with Y-27632, followed by collagenase IV and TrypLE Select dissociation whereupon small clumps were slow frozen using 90% Knockout serum replacement and 10% dimethyl sulfoxide. After thawing at 37 C, the clumps were cultured in medium supplemented with 10 microM Y-27632 for 1 day. Our results show that the use of Y-27632 significantly increases the survival of hES cells after thawing compared with that of the control group. Y-27632-treated freeze-thawed hES cells retain morphology, stable karyotype, expression of cell surface markers, and the potential to differentiate into derivatives of all three germ layers after long-term culture. We have concluded that conventional slow freezing with Y-27632 treatment is efficient and convenient for the cryopreservation of hES cells.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Canada.
authors.forename: Xiangyun
authors.initials: X
authors.lastname: Li
authors.forename: Guoliang
authors.initials: G
authors.lastname: Meng
authors.forename: Roman
authors.initials: R
authors.lastname: Krawetz
authors.forename: Shiying
authors.initials: S
authors.lastname: Liu
authors.forename: Derrick E
authors.initials: DE
authors.lastname: Rancourt
mesh_terms: D000577:Amides / Q000494:pharmacology*; D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D011725:Pyridines / Q000494:pharmacology*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D000577:Amides; D000943:Antigens, Differentiation; D003451:Cryoprotective Agents; D004791:Enzyme Inhibitors; D011725:Pyridines; C108830:Y 27632; D054460:rho-Associated Kinases; D004121:Dimethyl Sulfoxide