title: Derivation and long-term culture of transgene-free human induced pluripotent stem cells on synthetic substrates.
abstract: We describe a platform to derive, culture, and differentiate genomically stable, transgene-free human induced pluripotent stem cells (iPSCs) on a fully synthetic polymer substrate made of a grafted zwitterionic hydrogel: poly2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl) ammonium hydroxide (PMEDSAH). Three independent transgene-free iPSC lines derived in these conditions demonstrated continuous self-renewal, genomic stability, and pluripotency in vitro and in vivo after up to 9 months of continuous in vitro culture on PMEDSAH-grafted plates. Together, these data demonstrate the strength this alternative platform offers to generate and maintain human iPSCs for regenerative medicine.
authors.affiliation: Department of Biologic and Materials Sciences, Biointerfaces Institute, and Department of Chemical Engineering, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Luis Gerardo
authors.initials: LG
authors.lastname: Villa-Diaz
authors.affiliation: Department of Biologic and Materials Sciences, Biointerfaces Institute, and Department of Chemical Engineering, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Jin Koo
authors.initials: JK
authors.lastname: Kim
authors.affiliation: Department of Biologic and Materials Sciences, Biointerfaces Institute, and Department of Chemical Engineering, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Joerg
authors.initials: J
authors.lastname: Lahann
authors.affiliation: Department of Biologic and Materials Sciences, Biointerfaces Institute, and Department of Chemical Engineering, University of Michigan, Ann Arbor, Michigan, USA paulk@umich.edu.
authors.forename: Paul H
authors.initials: PH
authors.lastname: Krebsbach
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008689:Methacrylates / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D044968:Regenerative Medicine; D013997:Time Factors; D019076:Transgenes
chemical_list: D020100:Hydrogels; D008689:Methacrylates; D000644:Quaternary Ammonium Compounds; C568649:poly(2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl)ammonium hydroxide)
keywords: Differentiation; Feeder-free culture; Induced pluripotent stem cells; Regenerative medicine; Stem cells; Synthetic coating; iPSC