Fig. 4.

Directed differentiation of iPS cells into disease-relevant cell types in vitro. (A–F) Directed differentiation of Fabry-iPS cells into cardiomyocytes. (A) Spontaneously beating cell clusters occurred after attachment of the EBs to gelatin-coated dishes. The photograph was printed from a video image showing a beating cell cluster. (B) Many cells in the cluster shown in A stained for cardiac cell markers actinin (red) and GATA-4 (green; nuclear localization). (C and D) Enlarged view of Fabry-iPS–derived cardiomyocytes expressing actinin (red) (C), Troponin T (red) and GATA-4 (green) (D). (E) Cardiomyocytes differentiated from Fabry-iPS cells exhibit accumulated Gb3 in the cytoplasm. (Upper) Gb3 staining (red). (Lower) GATA-4 staining (green, nuclear localization). (F) Effect of diltiazem, an L-type calcium channel blocker on the frequency of beating of Fabry-iPS–derived cardiomyocytes. Data are presented as mean ± SD (n = 3). (G) Differentiation to neural stem cells from GLD-iPS cells. After enrichment of neural stem cells, almost all of the differentiated cells were strongly positive for nestin. [Scale bars: 20 μm (C–E); 200 μm (B and G).]