期刊
STEM CELLS
卷 34, 期 11, 页码 2670-2680出版社
WILEY
DOI: 10.1002/stem.2466
关键词
Human induced pluripotent stem cells; Cardiomyocyte subtype; Electrophysiology; Pacemaker
资金
- California Institute of Regenerative Medicine (CIRM) Basic Biology Grant [RB4-05764]
- CIRM Postdoctoral Training Fellowship [TG2-01163]
- National Institute of Health T32 Training Program in Basic and Translational Cardiovascular Science [T32HL086350]
- NIH [R01 HL085727, R01 HL085844, S10 RR033106]
- VA Merit Review Grant [I01 BX000576]
- National Science Foundation (NSF) [1264776]
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1264776] Funding Source: National Science Foundation
Insights into the expression of pacemaker-specific markers in human induced pluripotent stem cell (hiPSC)-derived cardiomyocyte subtypes can facilitate the enrichment and track differentiation and maturation of hiPSC-derived pacemaker-like cardiomyocytes. To date, no study has directly assessed gene expression in each pacemaker-, atria-, and ventricular-like cardiomyocyte subtype derived from hiPSCs since currently the subtypes of these immature cardiomyocytes can only be identified by action potential profiles. Traditional acquisition of action potentials using patch-clamp recordings renders the cells unviable for subsequent analysis. We circumvented these issues by acquiring the action potential profile of a single cell optically followed by assessment of protein expression through immunostaining in that same cell. Our same-single-cell analysis for the first time revealed expression of proposed pacemaker-specific markers-hyperpolarization-activated cyclic nucleotide-modulated (HCN) 4 channel and Islet (Isl) 1-at the protein level in all three hiPSC-derived cardiomyocyte subtypes. HCN4 expression was found to be higher in pacemaker-like hiPSC-derived cardiomyocytes than atrial-and ventricular-like subtypes but its downregulation over time in all subtypes diminished the differences. Isl1 expression in pacemaker-like hiPSC-derived cardiomyocytes was initially not statistically different than the contractile subtypes but did become statistically higher than ventricular-like cells with time. Our observations suggest that although HCN4 and Isl1 are differentially expressed in hiPSC-derived pacemaker-like relative to ventricular-like cardiomyocytes, these markers alone are insufficient in identifying hiPSC-derived pacemaker-like cardiomyocytes.
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