Table 1.

Related research.

Author	Published year	Organoid induction designs	Organoid type	Induction condition factors	Culture medium	SFEBq procedure
Approach 1: Stepwise, direct organization of region-specific or population-enriched organoids
Lakshmi Subramanian	2017	Neural tissue samples between GW8 and GW10; coronal vibratome sections were transferred to culture medium.	Forebrain	Wnt inhibitor; TGF-β inhibitor; high O2 penetration; concentration of matrigel	Cortical slice culture medium	No
Marina Bershteyn	2017	hIPSCs were cultured in using cortical differentiation medium.	Cerebral organoids	Rho kinase inhibitor; Wnt inhibitor; TGF-β inhibitor	Cortical differentiation medium; N2 culture medium	No
Adrian Ranga	2016	ESCs were cultured in high-throughput combinatorial screening of 3D microenvironments; stepwise induced.	Cyst-like structure neural tube with apical-basal polarity	RA; sonic hedgehog; synthetic nondegradable materials	Neurobasal medium	No
Lixiong Gao	2016	hESCs were cultured with SFEBq procedure; H1 cells were selected to induce organoids.	Neural retinal tissue	Wnt signal inhibitor; SAG; CHIR99021; N2; retinoic acid; high oxygen concentrate (40%)	N2 culture medium	Yes (3 days)
Hideya Sakaguchi	2015	hESCs were cultured in SFEBq culture for 73–84 days and dissociated with neural tissue dissociation kit and then cultured in neurobasal medium.	Hippocampal tissue	Wnt inhibitor; TGF-β inhibitor; N2 supplement; chemically defined lipid; neurobasal medium; bone morphogenetic protein; Wnt	N2; neurobasal medium	Yes (3 days)
Anca M. Paşca	2015	hCSs were dissociated to culture to induce organoids.	Cortical tissue with functional neural network	BMP inhibitor and TGF-β inhibitor; NT3; BDNF; FGF2; EGF; neurobasal; dorsomorphin; SB-431542	Neural medium	No
Atsushi Kuwahara	2015	Elective NR differentiation from hESCs.	Neural retina	BMP4-inhibiting GSK3 and FGFR; Y-27632; gfCDM; CHIR99021; SU5402	N2; neural retina medium; retinosphere medium	No
Karl R. Koehler	2013	ESCs were dissociated to induce organoids.	Inner ear sensory epithelial tissue	BMP4; TGF-β inhibitor FGF2 and LDN; N2B27 medium; N2 medium; SU5402; SB-431542	N2B27 medium	Yes (3 days)
Taisuke Kadoshima	2013	Stepwise induction of neocortex with high numbers of pyramidal neurons.	Neocortex	Rho kinase inhibitor, TGF-β inhibitor, and Wnt inhibitor; B27, N2, and chemically defined lipid concentration; hedgehog signals	N2 and chemically defined lipid concentration	Yes (3 days)
Lucy A. Crompton	2013	Neurospheres were dissociated cholinergic neurons to culture in NEM.	Forebrain cholinergic neurons	Nodal/TGF-β signaling inhibitor, ROCK inhibitor Y-27632, FGF2, and EGF	Modified chemically defined media, NEM	Yes (4 days)
Yichen Shi	2012	Neuroepithelial cells were dissociated to induce organoids.	Cerebral cortex with projection neurons and neural networks	SB431542, FGF2, noggin, neurobasal, Y-27632	N2B27 (N3) medium; N2	No
Jessica Mariani	2012	Undifferentiated PGP1-1 and colonies were dissociated into single cells to induce organoids.	Early forebrain	N2 supplement, Y-27632, FGF2, Wnt inhibitor DKK1, BMP inhibitor BMPRIA-Fc, TGF-β/activin/nodal inhibitor SB431542	N2	Yes (3 days)
Teruko Danjo	2011	Foxg1: venus+ cells were sorted to stepwise, induce to ventral telencephalic tissues.	Ventral telencephalic tissues	BDNF, vibratome, Shh	Neuron culture medium; NT3; neurobasal/B27	Yes (3 days)
Mototsugu Eiraku	2008	ESCs were sorted to induce of polarized cortical neuroepithelia.	Cortical tissues	FGF, Wnt, and BMP; N2 medium, KSR	Cortical slice culture medium	No
Kiichi Watanabe	2005	Stepwise induction of neocortex with high number of telencephalic precursors.	Cortical organoids with high number of telencephalic precursors	Dkk1, Bf1. LeftyA, Wnt3at, Wnt, and nodal	Cortical slice culture medium	Yes (3 days)
Approach 2: Assemble and direct disctinct organ-specific progenitor cells or stem cells to form specific morphogenesis organ
Chikafumi Ozone	2016	Coculture specific ventral hypothalamic NE tissue and nonneural ectoderm formation.	Anterior pituitary	KSR; FGF2; BMP4	gfCDM	Yes (3 days)
Keiko Muguruma	2015	hESCs were cultured to form neural tube-like NE structures and dissociated with anti-KIRREL2 antibody and then cocultured with RL tissues to generate Purkinje cells.	Polarized cerebellar tissue and Purkinje cells	SDF1 and FGF19; FGF2	gfCDM; N2	Yes (3 days)
Andrea Meinhardt	2014	Assembled R1 mESCs, IB10 mESCs, and 46C in 3D matrigel matrix and cocultured to induce the formation of neuroepithelial cyst.	Neuroepithelial cyst	RA, SAG, cyclopamine	N2B27 medium	No
Hidetaka Suga	2011	Assembled nonneural ectoderm and hypothalamic neuroectoderm cells in three-dimensional culture and cocultured to induce of adenohypophysis like tissue.	Adenohypophysis	SAG, DAPT, BIO, Wnt4, and Wnt5, FGF8, Nodal, IWP2, FGF10	CDM medium; DAPT-free medium	Yes (10 days)
Mototsugu Eiraku	2011	Assembled neuroectodermal epithelium tissue and nonretinal neuroectodermal epithelium in three-dimensional culture and cocultured to induce of optic cup.	Optic cup	40%-O2/5%-CO2, N2, RA, CUY21 generator	N2	Yes (3 days)
Xue-Jun Li	2008	Cultured neuroepithelial cells to generate motoneurons and then coculture of motoneurons and myocytes.	Ventral spinal tissue	RA and Shh, BDNF, GDNF, IGF1, Shh, B27	Neural differentiation medium; N2B27	No
Approach 2: 2.3 Assemble embryoid bodies for induction of multilayer organoids
Yun Li	2017	Differentiation of hESCs to EBs. EBs were embedded in droplets of matrigel. Embedded EBs were subsequently cultured to induce cerebral organoids.	Cerebral organoids	bFGF; ROCK inhibitor; orbital shaker; WIBR3; BDNF	Neurobasal; N2	Yes (6 days)
Madeline A. Lancaster	2013	Assembled EBs in the droplets of matrigel and cocultured to induce the formation of cerebral organoids.	Cerebral organoids	Retinoic acid, protein kinase (ROCK) inhibitor, neurobasal, N2, insulin, B27	N2B27	Yes (6 days)