Figure 6.

INS⁺ENTPD3⁺ cells are present in patient-derived seBC. A: Schematic representation of stepwise differentiation of iPSC clusters toward β-like cells in suspension. B: Representative flow cytometry analysis of iPSC differentiation at definitive endoderm (DE), pancreatic endoderm (PE), imBC, and seBC for specific lineage markers (n = 2 independent differentiation experiments). C: Immunofluorescence staining of seBC clusters derived from CTRL-iPSC, T1D-iPSC, and iPSC containing a GFP reporter under the contorl of the NKX6.1 promoter (scale bar represents 20 μm) (n = 5 independent differentiation experiments). D: Schematic representation of pINS⁺ENTPD3^+/− cells sorted from iPSC-derived seBC clusters reaggregated in the presence of support cells, HUVEC, and mesenchymal stem cells (MSC) for 48 h. E: Representative perifusion analysis of INS⁺ENTPD3⁻ and INS⁺ENTPD3⁺ clusters; 35–45 clusters were analyzed per condition, and data are presented as % of total insulin in cluster pellet recovered. EE, early endocrine.