Figure 3

Characterization of the pancreatic stem cell line D#22. A. RT-PCR analysis of expression of an ES cell marker (Nanog), pancreatic progenitor markers (Pdx1), and mature pancreatic markers (insulin-2, glucagon, amylase, and somatostatin) in D#22 cells. Pancreatic cells and iPS cells were used as controls. The primers used for RT-PCR are shown in Table  1. B. Quantitative RT-PCR analysis of the Ngn3 and NeuroD genes in iPS, D#22, and pancreatic cells. The data are expressed as the Ngn3 and NeuroD to β-actin ratio, with the expression in D#22 cells arbitrarily set at 1.0. C. Morphology of D#22 stage 5 cells and iPS stage 5 cells. Scale bars = 200 μm. D. Quantitative RT-PCR analysis of the insulin-2 gene in D#22 stage 5 cells and iPS stage 5 cells. Differentiated cells derived from D#22 cells by stages 3–5 or iPS cells by stages 1–5 were analyzed using quantitative RT-PCR. The data are expressed as the insulin-2 to β-actin ratio, with the expression in iPS stage 5 cells arbitrarily set at 1.0. E. RT-PCR testing for pancreatic markers of maturity (glucagon, amylase, and somatostatin) in D#22 stage 5 and iPS stage 5 cells. Pancreatic cells were used as a control. The primers used for RT-PCR are shown in Table  1.