Use of Adeno-Associated Virus to Enrich Cardiomyocytes Derived from Human Stem Cells

Hum Gene Ther Clin Dev. 2015 Sep;26(3):194-201. doi: 10.1089/humc.2015.052. Epub 2015 Aug 7.

Abstract

Cardiomyocytes derived from human induced pluripotent stem cells (iPSCs) show great promise as autologous donor cells to treat heart disease. A major technical obstacle to this approach is that available induction methods often produce heterogeneous cell population with low percentage of cardiomyocytes. Here we describe a cardiac enrichment approach using nonintegrating adeno-associated virus (AAV). We first examined several AAV serotypes for their ability to selectively transduce iPSC-derived cardiomyocytes. Results showed that AAV1 demonstrated the highest in vitro transduction efficiency among seven widely used serotypes. Next, differentiated iPSC derivatives were transduced with drug-selectable AAV1 expressing neomycin resistance gene. Selection with G418 enriched the cardiac cell fraction from 27% to 57% in 2 weeks. Compared with other enrichment strategies such as integrative genetic selection, mitochondria labeling, or surface marker cell sorting, this simple AAV method described herein bypasses antibody or dye labeling. These findings provide proof of concept for large-scale cardiomyocyte enrichment by exploiting AAV's intrinsic tissue tropism.

Publication types

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

MeSH terms

  • Cell Differentiation* / genetics
  • Cell Line
  • Dependovirus / classification
  • Dependovirus / genetics*
  • Gene Expression
  • Gene Transfer Techniques
  • Genes, Reporter
  • Genetic Vectors / genetics*
  • Humans
  • Immunohistochemistry
  • Immunophenotyping
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism*
  • Serogroup
  • Stem Cells / cytology*
  • Stem Cells / metabolism*
  • Transduction, Genetic
  • Viral Tropism