FIG. 7.

In vitro differentiation of iPS(IMR90)-4 cells. (A) Embryoid bodies (EBs) were derived from iPS(IMR90)-4 cells that had previously been cultured in 0.5% NFC hydrogel for 8 days followed by a 24-h-treatment with 300 μg cellulase/mg cellulose at 37°C. Cells dissociated from 1-week-old EBs were cultured on gelatin-coated dishes for 1 week and were then detected to express three germ layer markers, β-tubulin type III (b-tub), muscle actin (MA), and α-fetoprotein (AFP). Scale bars=100 μm. (B) Real-time quantitative RT-PCR analyses of PAX6, CDX2, and BRACHYURY mRNA expression in 4-week-old EBs derived from iPS(IMR90)-4 cells that had previously been cultured in 0.5 wt.% NFC hydrogel for 9 days. Relative mRNA expression was normalized to the control gene RPLP0, and fold inductions were calculated with reference to the cells cultured on the standard Matrigel platform (M ctrl). n=3 biological samples. Error bars are SD. Color images available online at www.liebertpub.com/scd