. Author manuscript; available in PMC: 2016 Aug 1.

Published in final edited form as: Trends Cardiovasc Med. 2013 Sep 17;24(1):32–44. doi: 10.1016/j.tcm.2013.06.002

Table 1.

Overview of the most commonly used PSC cardiac differentiation protocols, including monolayer, EB, and “co- culture” strategies. Time points at which various cytokines and small molecule inhibitors are added are denoted for each protocol along with the reported efficiencies for these protocols.

	Format		Time Points (Days)										Reported Efficiency	Refs
Chemically Defined	Monolayer (2D)	Basic Monolayer	D-2	D-1	D0	D1	D3	D4	D5	D7	D8	D14	≤90%	Paige et al., 2010
		Basic Monolayer		100ng/ml WNT3A^*	100ng/ml Activin A	10ng/ml BMP-4			200ng/ml DKK-1^*				≤90%	Paige et al., 2010
		Matrix Sandwich	D-2	D-1	D0	D1	D3	D4	D5	D7	D8	D14	≤98%	Zhang et al., 2012
		Matrix Sandwich	Matrigel Overlay		Matrigel Overlay 100ng/ml Activin A	10ng/ml BMP-4 5ng/ml bFGF							≤98%	Zhang et al., 2012
		Small Molecules	D-2	D-1	D0	D1	D3	D4	D5	D7	D8	D14	≤98%	Lian et al., 2012a
		Small Molecules			12 M CHIR99021 µg/ml Insulin^*		5 µM IWP-2 or IWP-4						≤98%	Lian et al., 2012a
	EB (3D)	Staged, defined	D-2	D-1	D0	D1	D3	D4	D5	D7	D8	D14	30–60%	Kattman et al., 2011
					0.4ng/ml BMP-4	0–12ng/ml Activin A 2.5–20 ng/ml BMP-4 5ng/ml bFGF			150ng/ml DKK-1 10ng/mlVEGF		10ng/ml VEGF 5ng/ml bFGF
					0.4ng/ml BMP-4			5.4 M SB-431542^* 0.6µM Dorsomorphin^*	150ng/ml DKK-1 10ng/mlVEGF		10ng/ml VEGF 5ng/ml bFGF
Chemically Undefined		Undefined	D-2	D-1	D0	D1	D3	D4	D5	D7	D8	D14	0.1–10%	Kehat et al., 2001
		Undefined			5–20% FBS or KSR								0.1–10%	Kehat et al., 2001
	Co-culture(2D)		D-2	D-1	D0	D1	D3	D4	D5	D7	D8	D14	≤25%	Mummery et al, 2007
	Co-culture(2D)				END2 conditioned medium								≤25%	Mummery et al, 2007

D2 and D1 refer to days prior to differentiation. D0-D14 refer to days at time of differentiation.

FBS: Fetal Bovine Serum. KSR: Knockout Serum replacement.

Refers to reported modifications of the original protocols aimed to enhance the cardiogenic potential of PSC.