Derivation and long-term culture of transgene-free human induced pluripotent stem cells on synthetic substrates

Stem Cells Transl Med. 2014 Dec;3(12):1410-7. doi: 10.5966/sctm.2014-0087. Epub 2014 Oct 13.

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.

Keywords: Differentiation; Feeder-free culture; Induced pluripotent stem cells; Regenerative medicine; Stem cells; Synthetic coating; iPSC.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Culture Techniques / methods*
  • Cells, Cultured
  • Humans
  • Hydrogels / chemistry*
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Methacrylates / chemistry*
  • Quaternary Ammonium Compounds / chemistry*
  • Regenerative Medicine
  • Time Factors
  • Transgenes*

Substances

  • Hydrogels
  • Methacrylates
  • Quaternary Ammonium Compounds
  • poly(2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl)ammonium hydroxide)