Design of a Vitronectin-Based Recombinant Protein as a Defined Substrate for Differentiation of Human Pluripotent Stem Cells into Hepatocyte-Like Cells
Fig 3
Maintenance of hPSCs on vitronectin variant-coated surfaces.
(A) Immunocytochemistry of NANOG, SOX17, POU5F1, and T (Brachyury) in K3 hiPS cells cultured on recombinant human vitronectin (hVTN)-, R-Fc- or NC-Fc-coated dishes for 5 days. Nuclei were counterstained with DAPI. Scale bar indicates 100 μm. (B) Differentiation potential of hiPS cells cultured on R-Fc or NC-Fc into the ectodermal (SOX1), mesodermal (T), and endodermal (SOX17) lineages was examined. 201B6 cells were maintained on Matrigel, R-Fc or NC-Fc for 7 passages and transferred on Matrigel-coated surface to induce differentiation. (C) Characterization of teratomas formed by human iPS cells cultured on the R-Fc-coated surface. Teratomas generated from three different human iPS cell lines (253G1, 454E2, and TIG120-4f1) contained tissues derived from all three germ layers. Typical tissues are shown. Ectoderm: mature and immature central nervous systems (CNS), and optic nerve; Mesoderm: cartilage, kidney, striated muscle, and vasculature; Endoderm: intestine. Scale bar indicates 100 μm.