title: 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.
authors.affiliation: 1 Department of Physiology and Pharmacology, School of Medicine, Wake Forest University Health Sciences , Winston-Salem, North Carolina.|2 Department of Rehabilitation Medicine, University of Washington , Seattle, Washington.|3 Institute for Stem Cell and Regenerative Medicine, University of Washington , Seattle, Washington.
authors.forename: Xuan
authors.initials: X
authors.lastname: Guan
authors.affiliation: 4 Fred Hutchinson Cancer Research Center , Seattle, Washington.|5 Department of Medicine, University of Washington , Seattle, Washington.
authors.forename: Zejing
authors.initials: Z
authors.lastname: Wang
authors.affiliation: 2 Department of Rehabilitation Medicine, University of Washington , Seattle, Washington.|3 Institute for Stem Cell and Regenerative Medicine, University of Washington , Seattle, Washington.
authors.forename: Stefan
authors.initials: S
authors.lastname: Czerniecki
authors.affiliation: 2 Department of Rehabilitation Medicine, University of Washington , Seattle, Washington.|3 Institute for Stem Cell and Regenerative Medicine, University of Washington , Seattle, Washington.
authors.forename: David
authors.initials: D
authors.lastname: Mack
authors.affiliation: 6 INSERM UMR 1089 IRT 1, Institut de Recherche Thérapeutique, Université de Nantes , Nantes, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: François
authors.affiliation: 6 INSERM UMR 1089 IRT 1, Institut de Recherche Thérapeutique, Université de Nantes , Nantes, France.
authors.forename: Veronique
authors.initials: V
authors.lastname: Blouin
authors.affiliation: 6 INSERM UMR 1089 IRT 1, Institut de Recherche Thérapeutique, Université de Nantes , Nantes, France.|7 Department of Molecular Genetics & Microbiology, College of Medicine, University of Florida, Gainesville, Florida.
authors.forename: Philippe
authors.initials: P
authors.lastname: Moullier
authors.affiliation: 2 Department of Rehabilitation Medicine, University of Washington , Seattle, Washington.|3 Institute for Stem Cell and Regenerative Medicine, University of Washington , Seattle, Washington.
authors.forename: Martin K
authors.initials: MK
authors.lastname: Childers
mesh_terms: D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D000229:Dependovirus / Q000145:classification / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism*; D065288:Serogroup; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014161:Transduction, Genetic; D056189:Viral Tropism