Figure 2. Current Methods for Cardiac Differentiation of Human PSCs.
The three major approaches for differentiation of human PSCs to CMs are summarized: embryoid body (EB), monolayer culture, and inductive co-culture. Methods for forming EBs range from a simple enzymatic partial dissociation of hPSC colonies to various methods to more precisely control EB cell number and EB size using microwells with forced aggregation (centrifugation), microwells to first expand hPSC colonies to a defined size, and micropatterned hPSC colonies of defined sizes. Alternatively, propagating hPSCs as monolayers on Matrigel with defined media can be used for cardiogenesis. For both EBs and monolayer approaches, stage-specific application of key growth factors (GFs) in defined media are required for optimal cardiogenesis, although some protocols use fetal bovine serum (FBS) or small molecules to induce cardiogenesis (see text for details of specific protocols). Co-culture of mechanically passaged hPSCs with visceral endodermal-like END2 cells takes advantage of cell signaling from END2 cells to promote cardiogenesis.