Pluripotency of clonal hESC reporter lines.(A–C) Outgrowths of differentiated cells from EBs derived from a clonal PGK-H2BmCherry hESC line showed coincident H2BmCherry (A) and DAPI (B) immunostaining visible in the merged fluorescent image (C). (D–I) Clonal PGK-H2BmCherry hESC-derived EBs developed endothelial (D–F) and neuronal (G–I) lineages. (J) Expression of endodermal genes alpha fetoprotein (AFP) and hPDX-1 by day 20 EBs of clonal PGK-H2BmCherry hESCs (lane 2) compared to undifferentiated cells (lane 1) and isolated human pancreatic islet tissue as positive control for hPDX-1 (lane 3) and day 20 parental H9 EBs as control for AFP (lane 3). (K) Pluripotency markers expressed by clonal PGK-H2BmCherry hESCs (lane 2) compared to no reverse transcriptase control (lane 1) and undifferentiated parental H9 cells (lane 3).
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