Neural lineage differentiation of human pluripotent stem cells: Advances in disease modeling

Table 1 Comparison of methods for neural induction from human pluripotent stem cells

Method	Neural induction outcomes	Significance	Ref.
Embryoid bodies; selected neural rosettes; 2D and 3D culture	Neural tube-like rosettes stained with Nestin, Musashi-1 and NCAM; positive neuronal markers MAP2 and TUJ1 expression	First study of neural progenitor differentiation from hPSCs	Zhang et al[22], 2001
SFEBq aggregate; sorting cells; 3D culture	Self-organized structure with four distinct zones: ventricular, early and late cortical-plate, and Cajal-Retzius cell zones	Pure 3D culture, provides the basis for the brain organoid method	Eiraku et al[23], 2008
Dual SMAD inhibition; 2D monolayer culture	Complete neural conversion of > 80% of hESCs	Mostly wild used method; also enables neural induction in 3D culture	Chambers et al[24], 2009
Dual SMAD inhibition combined with retinoid signaling; 2D monolayer culture	More than 95% of hPSCs were PAX6 and OTX1/2 cortical progenitor cells in 15 d	Improved the dual SMAD inhibition protocol and higher neural induction efficiency	Shi et al[62], 2012
Cortical organoid/spheroid; 3D culture	Form layered structure tissues partially mimicking human cerebral cortex	Mostly brain-like tissue with some functions	Lancaster et al[17], 2013; Paşca et al[26], 2015; Qian et al[27], 2016
Dual SMAD inhibition combined with Wnt, FGF and Notch inhibition	Generate functional cortical neuron in 16 d	Improved the dual SMAD inhibition protocol and accelerated neural induction	Qi et al[28]

EB: Embryoid body; FGF: Fibroblast growth factor; hPSCs: Human pluripotent stem cells; MAP2: Microtubule-associated protein 2; NCAM: Neural cell adhesion molecule; OTX: Orthodenticle homeobox; PAX6: Paired box 6; SFEBq: Serum-free floating culture of EB-like aggregates with quick aggregation.

Table 2 Comparison of methods for brain organoids generation from human pluripotent stem cells

Organoid type or brain region modeled	Method brief description	Model application	Ref.
EB-like aggregates; cerebral cortex	SFEBq, static suspension culture with cell sorting	Form self-organized structure mimicking the early cortiogenesis	Eiraku et al[23], 2008
Cerebral organoid; whole brain	Spinning bioreactor with Matrigel supporting	Form pyramidal identities with spatial separation mimicking the developing human brain at early stage; modeling microcephaly	Lancaster et al[17], 2013
Cortical neuroepithelium; cerebral cortex	Improved SFEBq, in 40% oxygen in Lumox plates	Inside-out layer pattern for human cortex	Kadoshima et al[86], 2013
Cortical spheroid; cerebral cortex	Static suspension culture with FGF-2 and EGF	Generated laminated cerebral cortex-like structure with some functions	Paşca et al[26], 2015
Cerebellar-plate-like neuroepithelium; cerebellum	Static suspension culture with FGF-19 and SDF-1	Mimicking the early development of human cerebellum	Muguruma et al[129], 2015
Telencephalic organoids; forebrain	Static suspension culture after neural rosettes isolation manually	Modeling autism spectrum disorder	Mariani et al[130], 2015
Dorsomedial telencephalic-like tissue; hippocampus	Improved SFEBq, in 40% oxygen	Modeling the development of human hippocampus	Sakaguchi et al[107], 2015
Forebrain organoids; cerebral cortex	Miniaturized spinning bioreactor	Zika virus exposure	Qian et al[27], 2016
Midbrain organoids; midbrain	Miniaturized spinning bioreactor	Midbrain organoids contained TH+ cells	Qian et al[27], 2016
Hypothalamic organoids; hypothalamus	Miniaturized spinning bioreactor	Modeling early hypothalamus development	Qian et al[27], 2016
Midbrain organoids; midbrain	Static suspension culture on orbital shaker	Midbrain produced neuromelanin and dopamine	Jo et al[131], 2016
Pituitary organoid; anterior pituitary	Improved SFEBq	Formed pituitary placode with pituitary hormone-producing cells	Ozone et al[132], 2016
Cerebral organoid; cerebral cortex	Microfilament-engineered organoids under agitation	Formed polarized cortical plate and radial units	Lancaster et al[133], 2017
Cerebral organoid; whole brain	Spinning bioreactor with Matrigel supporting	Brain organoids formed spontaneously active neuronal networks	Quadrato et al[134], 2017
Brain assembloids; assembly dorsal and ventral forebrain organoids	Static suspension culture	Modelling migration of human interneurons and their functional integration into microcircuits using healthy and timothy syndrome cell line	Birey et al[99], 2017
Fused cerebral organoids; assembly dorsal and ventral forebrain organoids	Static suspension culture with Matrigel supporting on orbital shaker	Modelling migration of human interneurons in cerebral cortex	Birey et al[99], 2017
Fused cortical organoids and MGE organoids	Static suspension culture on orbital shaker	Modelling migration of human interneurons	Xiang et al[101], 2017
Neoplastic cerebral organoid	Static suspension culture with Matrigel supporting on orbital shaker	Modelling brain tumorigenesis	Bian et al[135], 2018
Granted brain organoids in mouse	Spinning bioreactor	Formed functional networks and blood vessels in the grafts	Mansour et al[136], 2018
Cortical spheroid	Static suspension culture	Modelling Alzheimer’s disease	Yan et al[87], 2018
Cerebral organoids	Static suspension culture with Geltrex supporting on orbital shaker	Modelling Alzheimer’s disease	Gonzalez et al[93], 2018
Neuromuscular organoid	Static suspension culture supporting on orbital shaker	Formed functional neuromuscular junctions and modelling myasthenia gravis	Faustion Martins et al[137], 2020
Section spherical organoid	Manually slicing forebrain organoids	Sliced organoids exhibited separated upper and deep cortical layer	Qian et al[90], 2020
Cortico-motor assembloids; assembly cortical spheroids, spinal spheroids, and skeletal muscle spheroids	Static suspension culture	Modeling cortical-motor circuits	Andersen et al[18], 2020
Cortico-striatal assembloids; assembly cortical spheroids and striatal spheroids	Static suspension culture	Modeling cortical-striatal circuits and 22q13.3 deletion syndrome	Miura et al[102], 2020
Air-liquid interface cerebral organoids	Slicing mature organoids and cultured in air-liquid interface not completely submerged in liquid	Formed network with functional output	Giandomenico et al[138], 2019

ASD: Autism spectrum disorder; EGF: Epidermal growth factor; FGF2: Fibroblast growth factor 2; hPSCs: Human pluripotent stem cells; SDF: Stromal cell-derived factor; SFEBq: Serum-free floating culture of EB-like aggregates with quick aggregation.