Generation of electrophysiologically functional cardiomyocytes from mouse induced pluripotent stem cells

Stem Cell Res. 2016 Mar;16(2):522-30. doi: 10.1016/j.scr.2016.02.032. Epub 2016 Feb 23.

Abstract

Induced pluripotent stem (iPS) cells can efficiently differentiate into the three germ layers similar to those formed by differentiated embryonic stem (ES) cells. This provides a new source of cells in which to establish preclinical allogeneic transplantation models. Our iPS cells were generated from mouse embryonic fibroblasts (MEFs) transfected with the Yamanaka factors, the four transcription factors (Oct4, Sox2, Klf4 and c-Myc), without antibiotic selection or MEF feeders. After the formation of embryoid bodies (EBs), iPS cells spontaneously differentiated into Flk1-positive cardiac progenitors and cardiomyocytes expressing cardiac-specific markers such as alpha sarcomeric actinin (α-actinin), cardiac alpha myosin heavy chain (α-MHC), cardiac troponin T (cTnT), and connexin 43 (CX43), as well as cardiac transcription factors Nk2 homebox 5 (Nkx2.5) and gata binding protein 4 (gata4). The electrophysiological activity of iPS cell-derived cardiomyocytes (iPS-CMs) was detected in beating cell clusters with optical mapping and RH237 a voltage-sensitive dye, and in single contracting cells with patch-clamp technology. Incompletely differentiated iPS cells formed teratomas when transplanted into a severe combined immunodeficiency (SCID) mouse model of myocardial infarction. Our results show that somatic cells can be reprogrammed into pluripotent stem cells, which in turn spontaneously differentiate into electrophysiologically functional mature cardiomyocytes expressing cardiac-specific makers, and that these cells can potentially be used to repair myocardial infarction (MI) in the future.

Publication types

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

MeSH terms

  • Animals
  • Cell Differentiation
  • Cell Line
  • Cellular Reprogramming
  • Connexin 43 / metabolism
  • Disease Models, Animal
  • Fibroblasts / cytology
  • Flow Cytometry
  • Induced Pluripotent Stem Cells / cytology*
  • Mice
  • Mice, Inbred C57BL
  • Mice, SCID
  • Microscopy, Fluorescence
  • Myocardial Infarction / therapy
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / transplantation
  • Myosin Heavy Chains / metabolism
  • Patch-Clamp Techniques
  • Teratoma / metabolism
  • Teratoma / pathology
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transplantation, Homologous
  • Troponin T / metabolism

Substances

  • Connexin 43
  • Transcription Factors
  • Troponin T
  • Myosin Heavy Chains