Generation of pig iPS cells: A model for cell therapy

Núria Montserrat, Elena Garreta Bahima, Laura Batlle, Sophia Häfner, Alexandre Miguel Cavaco Rodrigues, Federico González, Juan Carlos Izpisua Belmonte

Research output: Contribution to journalArticlepeer-review

73 Scopus citations

Abstract

Reprogramming of pig somatic cells to induced pluripotent stem cells provides a tremendous advance in the field of regenerative medicine since the pig represents an ideal large animal model for the preclinical testing of emerging cell therapies. However, the current generation of pig-induced pluripotent stem cells (piPSCs) require the use of time-consuming and laborious retroviral or lentiviral transduction approaches, in order to ectopically express the pluripotency-associated transcription factors Oct4, Sox2, Klf4 and c-Myc, in the presence of feeder cells. Here, we describe a simple method to produce piPSC with a single transfection of a CAG-driven polycistronic plasmid expressing Oct4, Sox2, Klf4, c-Myc and a green fluorescent protein (GFP) reporter gene, in gelatine-coated plates, with or without feeder cells. In our system, the derivation of piPSCs from adult pig ear fibroblasts on a gelatine coating showed a higher efficiency and rate of reprogramming when compared with three consecutive retroviral transductions of a similar polycistronic construct. Our piPSCs expressed the classical embryonic stem cell markers, exhibit a stable karyotype and formed teratomas. Moreover, we also developed a simple method to generate in vitro spontaneous beating cardiomiocyte-like cells from piPSCs. Overall, our preliminary results set the bases for the massive production of xeno-free and integration-free piPSCs and provide a powerful tool for the preclinical application of iPSC technology in a large animal setting.

Original languageEnglish (US)
Pages (from-to)121-130
Number of pages10
JournalJournal of Cardiovascular Translational Research
Volume4
Issue number2
DOIs
StatePublished - Jan 1 2011

Keywords

  • Cardiomyocyte differentition
  • Clinical translation
  • Embryonic stem cells
  • iPS cells

ASJC Scopus subject areas

  • Molecular Medicine
  • Genetics
  • Pharmaceutical Science
  • Cardiology and Cardiovascular Medicine
  • Genetics(clinical)

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