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For example, markers of undifferentiated cells such as Oct4, hTert (human telomerase reverse transcriptase) and Dusp6 (dual specificity phosphatase 6) were shown to decrease reliably during early differentiation of hESCs, while Brachyury and GATA-4, markers of mesoderm and early cardiogenesis, were found to be suitable to monitor cardiac differentiation.
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In a recent study, RMS was used to monitor the molecular changes in live stem cells during cardiac differentiation, and then to correlate these changes to gold-standard fluorescence staining for the cardiac phenotype [65].
Chan et al. showed that, in addition to promoting cardiac differentiation, Mesp1 promoted hematopoietic differentiation and skeletal myogenic differentiation of ESCs in different induction conditions.
In addition, treating iCPC with Wnt inhibitor IWP2 facilitated even more efficient cardiac differentiation.
Lower cardiac gene expressions and percentage of cardiac troponin-I positive cells at pH 6.8 indicated that cardiac differentiation of ESCs was significantly compromised.
Overall, cardiac differentiation 'preferred' ECM-rigid culture substrates, and beating-behavior 'preferred' Col-soft culture substrates.
Using our technologies, a biphasic effect of Notch activation on cardiac differentiation was found: early activation in undifferentiated human embryonic stem cells (hESCs) promotes ectodermal differentiation, activation in specified cardiovascular progenitor cells increases cardiac differentiation.
Cardiac differentiation was confirmed using an embryoid body protocol by electrophysiology studies that verified the formation of nodal, atrial and ventricular-type cardiac cells from these cells [10].
Figure 7 compares the time-course average Raman spectra of typical EBs, beating (successful cardiac differentiation) and non-beating (unsuccessful cardiac differentiation) with the Raman spectra of individual hESC-derived CMs and a non-CM.
Cardiac differentiation was detected as coordinated beating.
Chromatin modifying enzymes play a critical role in cardiac differentiation.
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