Derivation of cardiac progenitor cells from embryonic stem cells

J Vis Exp. 2015 Jan 12:(95):52047. doi: 10.3791/52047.

Abstract

Cardiac progenitor cells (CPCs) have the capacity to differentiate into cardiomyocytes, smooth muscle cells (SMC), and endothelial cells and hold great promise in cell therapy against heart disease. Among various methods to isolate CPCs, differentiation of embryonic stem cell (ESC) into CPCs attracts great attention in the field since ESCs can provide unlimited cell source. As a result, numerous strategies have been developed to derive CPCs from ESCs. In this protocol, differentiation and purification of embryonic CPCs from both mouse and human ESCs is described. Due to the difficulty of using cell surface markers to isolate embryonic CPCs, ESCs are engineered with fluorescent reporters activated by CPC-specific cre recombinase expression. Thus, CPCs can be enriched by fluorescence-activated cell sorting (FACS). This protocol illustrates procedures to form embryoid bodies (EBs) from ESCs for CPC specification and enrichment. The isolated CPCs can be subsequently cultured for cardiac lineage differentiation and other biological assays. This protocol is optimized for robust and efficient derivation of CPCs from both mouse and human ESCs.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Cell Differentiation / physiology
  • Cell Line
  • Cells, Cultured
  • Embryoid Bodies / cytology
  • Embryonic Stem Cells / cytology*
  • Endothelial Cells / cytology
  • Flow Cytometry / methods
  • Humans
  • Mice
  • Myocytes, Cardiac / cytology*
  • Myocytes, Smooth Muscle / cytology