Microfluidic three-dimensional cell culture of stem cells for high-throughput analysis

Table 1 Features of microfluidic techniques for stem cell engineering

Features of microfluidic technology	Solutions	Stem cell applications	Ref.
Cell or ECM patterning	Microchannel	Co-culture of hMSCs and hNSCs	Yang et al[34], 2015
	Microwell-patterned substrate	Co-culture of hESCs and fibroblasts	Khademhosseini et al[35], 2006
Chemical gradient	Multichannel array	Regulation of hematopoietic stem cell fates	Mahadik et al[26], 2014
	Overlapping gradients	Neuronal commitment of mouse ESCs	Cosson et al[37], 2013
	Sink and source channel with continuous flow	Chemotaxis of NSCs	Xu et al[38], 2013
Cell-cell or cell-ECM interaction	Microchannel-groove	Monitoring of differentiation and migration of NSCs derived from hESCs	Lee et al[39], 2013
Shear stress	Flow	Behavior observation of MSCs	Zheng et al[40], 2012
Droplet	Encapsulation and emulsion	Construction of ADSC microenvironment	Sakai et al[41], 2011
		Construction of pre-hatching embryo	Agarwal et al[42], 2013
Sorting and separation	Optical tweezer	Sorting of hESCs	Wang et al[43], 2013
	Electrical impedance flow	Identifying the differentiation of state of single cell	Song et al[44], 2013
	Multiple dielectrophoresis	Sorting of hMSCs	Song et al[45], 2015
Hydrodynamic trapping	Chamber array	EB-trap array	Suri, et al[46], 2013
Integration and automation	Multi-arrayed chips and integrated systems (e.g., liquid handler, cell chamber, imaging system, software)	Automatic culture of stem cell-derived dopaminergic neurons	Kane et al[47], 2019
		Investigation of dynamic changes of hematopoietic stem cell condition	Dettinger et al[48], 2018
		Clonal analysis of hESCs differentiation pattern	Sikorski et al[49], 2015
Mechanical and electrical actuators	Mechanical stretch	Stretch-activated stem cell differentiation	He et al[51], 2018
	Magnetic resonance	Quantification of metabolic flux in leukemia stem cells	Jeong et al[52], 2017

ECMs: Extracellular matrixes; hMSC: Human mesenchymal stem cell; hESC: Human embryonic stem cell; NSC: Neural stem cell; ADSC: Adipose-derived stem cell; EB: Embryoid body.

Table 2 High-throughput screening analysis for stem cell engineering

Techniques for high-throughput screening	Advantages	Applications	Cell types	Ref.
Microarray technique	Cell encapsulation in hydrogel-matrix spots; minimal consumption of cells and reagents	Toxicity and phenotypic screening of NPCs	NPCs	Nierode et al[60], 2016
		Studying of the expansion of mouse ESC	ESCs	Fernandes et al[62], 2009
Microencapsulation using microfluidic flow focusing	Multiple generation of 3D cells	Study of embryogenesis	iPSCs	Agarwal et al[42], 2013
	Encapsulation of cells and ECMs; controlled physicochemical properties of gel beads	Study of ESC expansion	mESCs	Allazetta et al[63], 2013
		Co-culture of pancreatic islets and hMSCs	hMSCs	Headen et al[64], 2014
Silicon stamp for spotting protein	Control of gel stiffness for stem cell fates	Study of stem cell niche	hMSCs	Gobaa et al[65], 2011
ECM array	Mimicking of microenvironments	Study of stem cell niche	hASCs	Beachley et al[66], 2015
Microwell array using thermoformed cyclic olefin polymer	Round-bottom array, uniform size of well array	EB generation	mESCs	Vrij et al[67], 2016
Micro droplet array; hydrophobic-hydrophilic surface	Robotics-free sample handling; high throughput; low reagent consumption; high-content readouts	Screening of iPSC pluripotency and proliferation	iPSCs	Zhang et al[68], 2016
Serial dilution generator	Generation of different concentration, combination and temporal sequence of drugs	Effect of cytokine (Tgfβ3) on hBM-MSC	hBM-MSC	Occhetta et al[69], 2015
Microraft array	Mimicking of microenvironments and enhancement of contact	Study of stem cell fate by mimicking niche	Intestinal stem cells	Gracz et al[70], 2015
Micropattern-well hybrid	Compatibility of HCS	Screening of stem cell differentiation and drug screening	NPCs	Yu et al[71], 2018

ECMs: Extracellular matrixes; iPSC: Induced pluripotent stem cell; NPC: Neural progenitor cell; mESC: Mouse embryonic stem cell; hASC: Human adipose-derived stem cell; HCS: High-content screening.

Table 3 High-throughput screening-based three-dimensional organ- or organoid-on-a-chip

Organ type	Platform type	Cell type	Applications	Ref.
Liver	Organ-on-a-chip	Human iPSC -hepatocytes	Screening of hepatotoxic drugs	Ware et al[74], 2015
	Organ-on-a-chip	Hepatocyte-ESC line	Co-culture of multi-organ	Materne et al[75], 2015
	Organoid-on-a-chip	iPSCs	Tests of liver function	Schepers et al[76], 2016
Heart	Organ-on-a-chip	Human iPSC -cardiomyocytes	Screening of molecular inducer related to cardiac myocyte proliferation	Titmarsh et al[83], 2016
		Human iPSC - cardiomyocytes	Modeling of mitochondrial cardiomyopathy of Barth syndrome	Wang et al[84], 2014
	Organoid-on-a-chip	hPSCs	Study of cardiac maturation	Mills et al[86], 2017
Brain	Organ-on-a-hip	NPCs	Toxicity test with in vitro brain model of Alzheimer’s disease	Park et al[91], 2015
		NPCs	Calcium assay	Lai et al[92], 2012
		NPCs	In vitro test for Alzheimer’s disease	Yu et al[71], 2018
	Organoid-on-a-chip	Human iPSC	Model of neurodevelopment disorder by prenatal nicotine exposure	Wang et al[93], 2018
	Organ-on-a-chip	Human iPSC	High-throughput compound evaluation on three-dimensional networks of neurons and glia	Wevers et al[94], 2016
Intestine	Organoid-on-a-chip	Human iPSC	Study of response to exogenous stimuli	Workman et al[97], 2018
Kidney	Organoid-on-a-chip	Human iPSC	High-throughput screening format organoids for multidimensional phenotypic screening	Czerniecki et al[100], 2018

iPSC: Induced pluripotent stem cell; NPC: Neural progenitor cell; ESC: Embryonic stem cell.