Molecular beacon-based detection and isolation of working-type cardiomyocytes derived from human pluripotent stem cells

Biomaterials
Rajneesh JhaChunhui Xu

Abstract

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) provide a potential source of cells to repair injured ventricular myocardium. CM differentiation cultures contain non-cardiac cells and CMs of both nodal and working subtypes. Direct application of such cultures in clinical studies could induce arrhythmias; thus, further purification of working-type CMs from heterogeneous cultures is desirable. Here, we designed 10 molecular beacons (MBs) targeting NPPA mRNA, a marker associated with working-type CMs and highly up-regulated during differentiation. We examined these MBs by solution assays and established their specificity using NPPA-overexpressing CHO cells as well as hPSC-CMs. We selected one MB for subsequent CM subtype isolation using fluorescence-activated cell sorting because the signal-to-background ratio was the highest for this MB in solution assays and a linear correlation was observed between MB signals and the CM purity in differentiation cultures. Compared with cells with low MB signals, cells positively selected based on MB signal had higher expression levels of genes associated with working-type CMs and lower expression levels of genes associated with nodal-type CMs. Therefore, the MB-based method is cap...Continue Reading

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Citations

Oct 19, 2017·Stem Cell Research & Therapy·Patrick A SchweizerDierk Thomas
Jan 27, 2019·Expert Opinion on Biological Therapy·Wenbin LiangDarryl R Davis
Jun 4, 2020·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Jiandong PanHao Chen
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