Laser-Based Propagation of Human iPS and ES Cells Generates Reproducible Cultures with Enhanced Differentiation Potential
Figure 5
iPSCs propagated by laser-mediated passage differentiated more efficiently into cardiomyocytes. (a) Brightfield image of day 4 EBs generated from iPSC cultures (BIMR A) propagated by laser-mediated passage, trypsin dissociation, or collagenase treatment. Scale bar, 250 μm. (b) Size of EBs generated from iPSC cultures propagated by laser-mediated passage, trypsin dissociation, or collagenase treatment ( EBs per data point). Data are shown as scatter plot with red line indicating mean and CV shown in red text above each sample. The asterisks (*) indicate variances that are statistically significant when compared to laser using ANOVA, with considered significant. (c) Percentage of EBs containing contracting areas. Data are shown as mean + s.d. ( independent experiments containing 75 EBs/sample in each experiment). (d) QRT-PCR analysis of cardiomyocyte-associated gene expression in EBs generated using iPSC cultures propagated by laser-mediated passage, trypsin dissociation, or collagenase treatment. The asterisks (*) indicate values that are statistically significant as compared with EBs generated from collagenase passaged iPSC cultures. The data are presented as mean ± s.d. (). Statistical analysis was performed using -test with considered significant. (e) Expression of cardiomyocyte markers, -MHC and -actinin, in EBs generated from iPSC cultures propagated by laser-mediated passage or collagenase treatment on day 22 of cardiac differentiation. Hoechst was used as a nuclear counterstain. Scale bars, 250 μm.
