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