Our group has developed a defined culture system to direct the differentiation of hPSCs into a near-homogenous population of definitive endoderm (DE) cells that have the capacity to differentiate into hepatocytes and pancreatic progenitors (Brown et al., 2011; Cho et al., 2012; Rashid et al., 2010; Touboul et al., 2010; Vallier et al., 2009a; Yusa et al., 2011). Cells grown in these culture conditions successively express primitive streak markers (T and Mixl1), downregulate pluripotency markers (NANOG, SOX2, and POU5F1) and progressively upregulate definitive endoderm markers (CXCR4, FOXA2, GATA4, CERB, and SOX17) (Figures S1A–S1C available online). Flow cytometry analyses showed that 80% of the resulting DE population coexpresses CXCR4 and SOX17 (Figure S1D). Interestingly, the resulting population of DE cells is negative for genes marking the foregut (SOX2), the midgut/hindgut (CDX2), the pancreas (PDX1), the liver (AFP), and the lungs (HOXA1) (Figures S1E and S1F) This confirms that DE cells generated in vitro could correspond to early endoderm progenitor cells prior to anteroposterior patterning or organogenesis.
resulting DE population
coexpresses CXCR4 and SOX17
80%
%