Human C-kit+CD45- cardiac stem cells are heterogeneous and display both cardiac and endothelial commitment by single-cell qPCR analysis

Biochem Biophys Res Commun. 2014 Jan 3;443(1):234-8. doi: 10.1016/j.bbrc.2013.11.086. Epub 2013 Dec 2.

Abstract

C-kit expressing cardiac stem cells have been described as multipotent. We have previously identified human cardiac C-kit+CD45- cells, but only found evidence of endothelial commitment. A small cardiac committed subpopulation within the C-kit+CD45- population might however be present. To investigate this at single-cell level, right and left atrial biopsies were dissociated and analyzed by FACS. Only right atrial biopsies contained a clearly distinguishable C-kit+CD45- population, which was single-cell sorted for qPCR. A minor portion of the sorted cells (1.1%) expressed early cardiac gene NKX2.5 while most of the cells (81%) expressed late endothelial gene VWF. VWF- cells were analyzed for a wider panel of genes. One group of these cells expressed endothelial genes (FLK-1, CD31) while another group expressed late cardiac genes (TNNT2, ACTC1). In conclusion, human C-kit+CD45- cells were predominantly localized to the right atrium. While most of these cells expressed endothelial genes, a minor portion expressed cardiac genes.

Keywords: Atrium; Cardiac stem cells; FACS; Heart; Human; SEM; SP; Side Population; Single-cell qPCR; qPCR; quantitative real-time polymerase chain reaction; standard error of the mean.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / analysis
  • Biomarkers / metabolism
  • Cell Differentiation*
  • Cell Lineage
  • Cell Separation
  • Endothelium, Vascular / cytology*
  • Flow Cytometry
  • Heart Atria / cytology
  • Humans
  • Leukocyte Common Antigens / analysis
  • Leukocyte Common Antigens / genetics
  • Leukocyte Common Antigens / metabolism*
  • Myoblasts, Cardiac / cytology*
  • Myocardium / cytology*
  • Proto-Oncogene Proteins c-kit / analysis
  • Proto-Oncogene Proteins c-kit / genetics
  • Proto-Oncogene Proteins c-kit / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Single-Cell Analysis

Substances

  • Biomarkers
  • Proto-Oncogene Proteins c-kit
  • Leukocyte Common Antigens
  • PTPRC protein, human