Fig 2.

Feeder-free hESC culture in NPY medium. H9 hESCs were maintained in MEF-CM or UM containing either PBS (control; -), 0.5 μM scrambled control SC-10, 0.5 μM scrambled control SC-36, 0.5 μM NPY or 40 ng/ml bFGF for three passages. Peptides were added to fresh medium daily. (A) Representative morphology of H9 hESCs cultured under various medium conditions as indicated. Upper panels: phase contrast images. Bar = 500 μm or 1 μm (inset images). Lower panels: scanned images of 35 mm round culture dishes and inverted macroscopic images were acquired after ALP staining. Bar = 500 mm (inset images). (B) Real-time qRT-PCR analysis for the expression of OCT4, SOX2, hTERT and TDGF. H9 and HES-7 hESCs were cultured using the indicated media for one or two passages. The results are displayed as the relative mRNA level with the level detected in undifferentiated hESCs cultured in CM set to 1 and are presented as the mean ± S.E. **P < 0.01, *P < 0.05, by t-test. (C) FACS analysis of SSEA-4 and ALP expression on H9 hESCs. Representative plots following flow cytometry from three independent experiments are shown.