Inducing human induced pluripotent stem cell differentiation through embryoid bodies: A practical and stable approach

Table 1 Updated summary of the formation of human embryoid bodies

Target cells or organoids	Culture vessels	Cell separation (cell suspension)	Culture period(d)	Media and supplements for EB formation	Ref.
Human brain organoids	Micropillar array plates	Accutase (single-)	11	KSR, Y27632, NEAA, GlutaMAX, bFGF	Zhu et al[65], 2017
Midbrain-like organoids	Low adhesion 96-well V-bottom multi-well plates	NA (single-)	4	mTeSR1, Y27632, Matrigel	Jo et al[21], 2016
Corneal epithelial cells	Static suspension culture	NA (non-single-)	14	E6 medium, Y27632, IWP-2, bFGF	Martínez García de la Torreet al[66], 2017
Osteoblast-like cells		Collagenase type IV, trypsin–EDTA (non-single-)	6	Human ES medium, thiazovivin	Ochiai-Shinoet al[67], 2014
Cardiac cells		NA (non-single-)	6	DMEM/F-12, NEAA, GlutaMAX, FBS, DMSO	Hoque et al[68], 2018
Neurospheres		Accutase (single-)	5	KSR, Y27632, FGF-2, NEAA, L-Glutamine	Pauly et al[69], 2018;
Cardiomyocytes		Type IV collagenase (single-)	10	DMEM, FBS, NEAA, GlutaMAX	Chauveau et al[70], 2017;
Osteoprogenitors		Dispase (non-single-)	7	DMEM, FBS, Glutamax, ABAM	Roberts et al[71], 2017
Chondrocytes		NA (single-)	6	Aggrewell medium, TeSR-E8 medium	Nam et al[72], 2017
Hepatocytes		Trypsin and collagenase IV (non-single-)	5	Knockout DMEM, NEAA, L-glutamine, FBS	Tomotsune et al[17], 2016
Oligodendrocytes		Trypsin-EDTA (non-single-)	2	KnockOut-DMEM, KSR, NEAA, SB431542, glutamine, ITS, Dorsomorphin	Espinosa-Jeffreyet al[73], 2016
Hematopoietic stem cells		Dispase (non-single-)	7	KnockOut-DMEM, KSR, Y27632, GlutaMAX, NEAA	Phondeechareon et al[74], 2016
Neural precursor cells		EDTA (non-single-)	5	DMEM/F12, KSR, NEAA, SB431542, GlutaMAX, Recombinant Noggin	Plaisted et al[75], 2016
Mesenchymal stem cells (MSCs)		Collagenase type IV (non-single-)	10	DMEM/F12, KSR, MEF conditioned medium, NEAA, bFGF, glutamine	Tang et al[76], 2014
Chondrocyte-like cells	Non-adherent 96-well plate	Collagenase IV (non-single-)	7	DMEM F12, L-glutamine, FBS, KSR, NEAA	Suchorska et al[77], 2017
Chondrocyte-like cells	Low adhesion 96-well C-bottom multi-well plate	Trypsin-EDTA (single-)	1	DMEM/F12, Y27632, KSR, NEAA	Lach et al[78], 2018
Retinal tissue	96-well non-adherent U-bottom plate	Accutase (single-)	12	mTeSR1, Y27632	Hunt et al[79], 2017
Insulin-secreting cells	Agarose round-bottom microwell plate	Accutase (single-)	10	IMDM/F-12, FBS, insulin transferring selenium-A, monothioglycerol	Pettinato et al[52], 2014
Neural precursors	Static or dynamic suspension culture method (orbital shaker)	EDTA (non-single-)	6-9	mTeSR1, Y27632; special equipment: orbital shaker	Miranda et al[80], 2016
Macrophages	AggreWellTM, 96-well round-bottomed multi-well plate	Collagenase-dispase mixture (single-)	4	DMEM/F12, Y27632, KSR, L-glutamine	Mukherjee et al[81], 2018
Primordial germ cell-likecells	AggreWell microwells	Accutase (single-)	1	KnockOut DMEM, Y27632, KSR, NEAA, bovine pancreas insulin, LIF, TGF-β, CHIR99021, PD0325901, BIRB796, SP600125	Mitsunaga et al[41], 2017
Forebrain cortical cells		Collagenase (non-single-)	4	DMEM/F12, KSR, glutamine, NEAA	Muratore et al[49], 2014
Melanocytes	24-well ElplasiaTMmicrowell plates (round bottom)	Accutase (single-)	7	mTeSR1, Y27632	Liu et al[82], 2019

“NA” means enzyme deficiency or mechanical dissection of the iPSC clones. E6 medium: Essential 6 medium; IMDM/F-12: Iscove's Modified Dulbecco's Medium/Nutrient Mixture F-12; DMEM/F-12: Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12; mTeSR1: Standardized medium for the feeder-independent maintenance of hESCs and hiPSCs; TeSR-E8 medium: A feeder-free, animal component-free culture medium for hESCs and hiPSCs; MEF: Mouse embryonic fibroblast; FBS: Fetal bovine serum; LIF: Leukemia inhibitory factor; TGF-β: Transforming growth factor-beta; bFGF: Basic fibroblast growth factor; Y27632: Rho-associated kinas inhibitor; KSR: Knockout serum replacement; NEAA: Non-essential amino acids; DMSO: Dimethyl sulphoxide; IWP-2: Inhibitor of the WNT pathway; ITS: Insulin-Transferrin-Selenium; SB431542: Inhibitor of the activin type I receptor ALK4 and the nodal type I receptor ALK7; CHIR99021: Inhibitor of glycogen synthase kinase 3β (GSK3β); PD0325901: Inhibitor of the mitogen-activated protein kinase (MEK) pathway; BIRB796: Inhibitor of the mitogen-activated protein kinases (MAPK); SP600125: Inhibitor of c-Jun N-terminal kinase (JNK).