De Novo Human Cardiac Myocytes for Medical Research: Promises and Challenges

Table 1

Strategies to promote cardiomyocyte maturation.

Strategies	Approaches	Findings	References	Pros/cons
Prolonged cell culture	3-month culture	Increased and	[37]	Simple protocol but time-consuming
	30-day culture	Increased conduction velocity	[38]
	80–120-day culture	More adult-like morphology and subcellular structure	[39]
Electrical stimulation	3–6 Hz pacing in 3D cultured hESC-CMs	Adult-like structure, calcium handling and electrical properties	[40]	Requires special equipment and optimization of stimulation protocol
	1 Hz in monolayer culture	Prolonged action potential, but no effects on resting membrane potential	[41]
	2 Hz in 3D tissue	Increased contractile force	[42]
Mechanical stretch	Cyclic stretch	Increased cell size, better sarcomere structure	[43]	Requires special equipment and optimization of stimulation protocol
	Cyclic stretch	Cell elongation, increased connexin expression, more mature calcium handling	[44]
Adrenergic receptor stimulation	Phenylephrine, 48 h	Increased cell size, better sarcomere structure	[43]	Simple protocol, easy to apply
Thyroid hormone stimulation	mESC-CMs: T3 for 7 days	Adult-like action potential profile and calcium handing	[46]	Simple protocol, easy to apply
	hiPSC-CMs: T3 for 7 days	Increased contractile force and faster calcium transient kinetics	[47]
MicroRNA overexpression	miR-1	Hyperpolarized resting , faster Ca^2+ transient kinetics	[48]	Requires introduction of foreign substance into cells