Integrin Based Isolation Enables Purification of Murine Lineage Committed Cardiomyocytes

PloS One
Laura TarnawskiStefan Jovinge

Abstract

In contrast to mature cardiomyocytes which have limited regenerative capacity, pluripotent stem cells represent a promising source for the generation of new cardiomyocytes. The tendency of pluripotent stem cells to form teratomas and the heterogeneity from various differentiation stages and cardiomyocyte cell sub-types, however, are major obstacles to overcome before this type of therapy could be applied in a clinical setting. Thus, the identification of extracellular markers for specific cardiomyocyte progenitors and mature subpopulations is of particular importance. The delineation of cardiomyocyte surface marker patterns not only serves as a means to derive homogeneous cell populations by FACS, but is also an essential tool to understand cardiac development. By using single-cell expression profiling in early mouse embryonic hearts, we found that a combination of integrin alpha-1, alpha-5, alpha-6 and N-cadherin enables isolation of lineage committed murine cardiomyocytes. Additionally, we were able to separate trabecular cardiomyocytes from solid ventricular myocardium and atrial murine cells. These cells exhibit expected subtype specific phenotype confirmed by electrophysiological analysis. We show that integrin expression ...Continue Reading

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Methods Mentioned

BETA
fluorescence-activated cell sorting
FACS
transgenic
FCS
Assay
PCR
chip
flow cytometry

Software Mentioned

censReg
Graph Pad Prism
R
GenEx ( MultiD )
Fluidigm analysis
Fluidigm

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