Data Sheet 1_CD98hc marks a precursor of human stem cell-derived alpha cells primed for functional maturation.pdf

Introduction<p>Pluripotent stem cell-derived alpha cells (SC-α cells) represent a promising component of next-generation cell therapies designed to more fully recapitulate human islet function; however, heterogeneity and the presence of undesired side populations limit their functional maturity and therapeutic potential. Here, we identify the cell surface protein CD98hc (SLC3A2), that when highly expressed in precursor cells, is associated with differentiation toward a more transcriptionally and functionally mature population of human SC-α cells. </p>Methods<p>Directed differentiation was used to generate SC-α cells from pluripotent stem cells, which were assessed via single cell RNA sequencing, flow cytometry, and immunofluorescence imaging.

Following magnetic activated cell sorting (MACS) of CD98hc enriched precursors, SC-α cell function was assessed using glucose stimulated glucagon secretion (GSGS).</p>Results<p>We show that CD98hc is upregulated in SC-α cells expressing high levels of genes known to play important roles in α cell identity and function, and that CD98hc expression is elevated at the pre-α precursor stage relative to non–α cell populations.