Abstract
The use of human stem cell-derived cardiomyocytes to study atrial biology and disease has been restricted by the lack of a reliable method for stem cell-derived atrial cell labeling and purification. The goal of this study was to generate an atrial-specific reporter construct to identify and purify human stem cell-derived atrial-like cardiomyocytes. We have created a bacterial artificial chromosome (BAC) reporter construct in which fluorescence is driven by expression of the atrial-specific gene sarcolipin (SLN). When purified using flow cytometry, cells with high fluorescence specifically express atrial genes and display functional calcium handling and electrophysiological properties consistent with atrial cardiomyocytes. Our data indicate that SLN can be used as a marker to successfully monitor and isolate hiPSC-derived atrial-like cardiomyocytes. These purified cells may find many applications, including in the study of atrial-specific pathologies and chamber-specific lineage development.
Highlights
The ability to differentiate human pluripotent stem cells into cardiomyocytes is a promising strategy for understanding human cardiac biology and disease [1]
We have provided a proof-of-concept study to show that SLN expression can be used as a marker to successfully monitor and isolate hiPSC-derived atrial-like myocytes
SLN expression appears concurrent with the onset of beating, and continues for extended periods in culture, allowing for isolation of highly red fluorescent atrial-like cells at early or later time-points during differentiation
Summary
The ability to differentiate human pluripotent stem cells into cardiomyocytes is a promising strategy for understanding human cardiac biology and disease [1]. The expression of one gene, sarcolipin (SLN), an inhibitor of the sarcoplasmic reticulum Ca2+-ATPase (SERCA), is restricted to the atrial lineage in the developing mouse heart from the onset of its expression, and this pattern is conserved in other mammals including humans [8,9,10]. It is unknown if SLN expression can be used to discriminate human atrial cells in differentiating pluripotent stem cell cultures, and if derived SLN-expressing cells would resemble functional atrial cardiomyocytes.
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