Activin A Modulates CRIPTO-1/HNF4α⁺ Cells to Guide Cardiac Differentiation from Human Embryonic Stem Cells

Figure 3

ActA increased human ESC-CM differentiation efficiency. High doses of 50 and 100 ng/mL ActA promoted CM differentiation. (a) Brightfield images of EB morphology at days 5, 7, and 9 of differentiation of ActA treatment conditions (50 and 100 ng/mL) compared to the untreated cells (0 ng/mL ActA). (b) Proliferation was assessed using Ki-67, showing that 6.95% of the control cells and 8.94% of the ActA treated cells were in a proliferative stage. Flow cytometry analysis is a representative example of three independent experiments. (c) Quantification of contracting rate (beating frequency EBs per minute) and (d) percentage of contracting EBs at day 10 of differentiation. Efficiency is expressed as percentage of wells containing beating EBs to the total number of wells. (e) Gene expression analyses of late-stage CM genes (TNNT2, cMyHC) during 13 days of differentiation. Dashed lines show basal expression levels of undifferentiated ESCs. Each data point is represented as , normalized for the housekeeping genes GAPDH, HPRT, and RPL13a. Data are representative of three independent experiments and expressed as mean SEM. Significant differences are indicated as : versus control and + versus 50 ng/mL; : versus control; : ∗∗∗ versus control. Scale bar = 100 μm.

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