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Stem Cell Res. 2013 Nov;11(3):1335-47. doi: 10.1016/j.scr.2013.09.003. Epub 2013 Sep 18.

Myosin light chain 2-based selection of human iPSC-derived early ventricular cardiac myocytes.

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Department of Internal Medicine, Cardiovascular Medicine, Center for Arrhythmia Research, University of Michigan, Ann Arbor, MI 48108, USA; Université Pierre et Marie Curie, Paris, France.


Applications of human induced pluripotent stem cell derived-cardiac myocytes (hiPSC-CMs) would be strengthened by the ability to generate specific cardiac myocyte (CM) lineages. However, purification of lineage-specific hiPSC-CMs is limited by the lack of cell marking techniques. Here, we have developed an iPSC-CM marking system using recombinant adenoviral reporter constructs with atrial- or ventricular-specific myosin light chain-2 (MLC-2) promoters. MLC-2a and MLC-2v selected hiPSC-CMs were purified by fluorescence-activated cell sorting and their biochemical and electrophysiological phenotypes analyzed. We demonstrate that the phenotype of both populations remained stable in culture and they expressed the expected sarcomeric proteins, gap junction proteins and chamber-specific transcription factors. Compared to MLC-2a cells, MLC-2v selected CMs had larger action potential amplitudes and durations. In addition, by immunofluorescence, we showed that MLC-2 isoform expression can be used to enrich hiPSC-CM consistent with early atrial and ventricular myocyte lineages. However, only the ventricular myosin light chain-2 promoter was able to purify a highly homogeneous population of iPSC-CMs. Using this approach, it is now possible to develop ventricular-specific disease models using iPSC-CMs while atrial-specific iPSC-CM cultures may require additional chamber-specific markers.


AP; Ad; CM; ESC; FACS; GFP; MLC-2ap; MLC-2vp; action potential; adenovirus; cardiac myocyte; embryonic stem cell; fluorescence-activated cell sorting; green fluorescent protein; hiPSC; human induced pluripotent stem cell; myosin light chain-2 atrial promoter; myosin light chain-2 ventricular promoter

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