Table 1.

A comprehensive summary of 3D cell culture methods.

Method	Culture cell types	Viability	Proliferation	Differentiation	Time length	Spheroid diameter (μm)	Advantages	Disadvantages	Refs.
Liquid overlay	(1) MDPSCs; (2) ADSCs	Well-preserved		Osteogenic	-1 day	30-100	(1) Simple; (2) economical	Size of the spheroids cannot be controlled	[15–21]
Hanging drop culture	WJ-MSCs	Cell death rate was below 40%	Higher than 2D	Early osteogenic	Depend on cell types	Controllable	Uniformity	Volume cannot more than 30 μL	[2, 22–27]
Rotating bioreactor	BMSCs	High level of viability		Osteoblastic	1 day	100–200	(1) Simple; (2) efficient	(1) Shear stress; (2) equipment	[18, 28]
Magnetic suspension	NIH3T3	99% cell viability	Exponential growth		5 minutes		(1) Repeatable ability; (2) size stability.	Potential impact of nanoparticles is uncertain	[24, 29–34]
Chemical reagents culture	ADSCs	High level of viability	Suppressed	Osteogenesis	3-4 days	50-200	(1) Simple; (2) practical; (3) special equipment unnecessary	Potential impacts of reagent are uncertain	[6, 35]
Alginate-PEG gels	mMSCs	Improves the viability of cell	Reducing cellular apoptosis	Osteogenic			Biocompatibility high mechanical strength	Early complicate	[36–57]