Table 1 Experimental protocols for motor neuron induction from human embryonic and induced pluripotent stem cells
References | Starting cells | Neural induction | MN differentiation | MN maturation | Duration | ||||
|---|---|---|---|---|---|---|---|---|---|
Medium | Factors | Obtained cells | Medium | Factors | Medium | Factors | |||
Amoroso et al. [39] | ESCs and iPSCs | ESC mediuma | Y27632, bFGF, LDN193189, and SB435142 | Embryoid bodies | Neural induction mediumb | RA, ascorbic acid, BDNF, C25II modified Shh, SAG, HAG, or purmorphamine | Neurobasal medium | IGF1, GDNF, CNTF, and B27 | 20-30 days |
Boulting et al. [40] | ESCs and iPSCs | ESC medium with 10% KO replacement | bFGF and Y27632 | Embryoid bodies | Neural induction mediumb supplemented with bFGF | RA, ascorbic acid, db-cAMP, and HAG | Neurobasal medium | BDNF, GDNF, and CNTF | 29Â days |
Burkhardt et al. [41] | iPSCs | DMEM/F12 supplemented with N2, B27, and NEAA | Y27632, Dorsomorphin dihydrochloride | Embryoid bodies | DMEM/F12, GLUTAMAX, N2, B27 serum-free supplement, D-Glucose, and ascorbic acid | SAG, RA, and DAPT (only for 4Â days) | DMEM/F12, GLUTAMAX | CNTF, BDNF, and GDNF | 32Â days |
N2, B27 serum free supplement, D-Glucose, and ascorbic acid | |||||||||
Corti et al. [25] | iPSCs | DMEM/F12, NEAA, N2, and heparin | RA | Neural rosette | DMEM/F12, NEAA, N2, and heparin | RA and Shh | DMEM/F12, NEAA, N2, and heparin | BDNF, GDNF, and IGF1 | ~24Â days |
Hester et al. [42] | ESCs and iPSCs | DMEM/F12, N2, and 10% KO serum | - | Embryoid bodies then neural rosette | DMEM/F12 with N2 | RA, Shh, forskolin, B27, and adenovirus | - | - | ~40Â days |
Hu and Zhang [43] | ESCs | DMEM/F12, KO serum replacement, NEAA, L-Glu, and BME | - | Floating embryoid bodies | Neural differentiation medium (DMEM/F12, N2, NEAA, and heparin) | Shh or purmorphamine, RA, B27, cAMP, ascorbic acid, BDNF, GDNF, and IGF1 | Neural differentiation medium (DMEM/F12, N2, NEAA, and heparin) | cAMP, ascorbic acid, BDNF, GDNF, and IGF1 | ~40Â days |
Karumbayaram et al. [44] | iPSCs and ESCs | ESC medium without FGF2 | - | Embryoid bodies/neural rosette | ESC medium without FGF2 | RA and purmorphamine | DMEM/F12 and N2 | GDNF, BDNF, CNTF, Shh, and RA | 35-49 days |
Reinhardt et al. [29] | iPSCs and ESCs | ESC mediuma and then expansion medium (DMEM/F12, N2, B27, pen/strep, and L-Glu) | For induction: SB435142, Dorsomorphine, CHIR, and purmorphamine | Embryoid bodies (smNPC) | Expansion medium (DMEM/F12, N2, B27, pen/strep, and L-Glu) | Purmorphamine and RA | Expansion N2, B27 | BDNF, GDNF, db-cAMP, RA, and purmorphamine | >40Â days |
For expansion: ascorbic acid, CHIR99021, and purmorphamine | |||||||||
Sareen et al. [45] | iPSCs | Neural differentiation medium (DMEM, B27, vitamin A, and N2) | RA | Embryoid bodies | Neurobasal medium, B27, and N2 | RA and purmorphamine | DMEM/F12 and B27 | RA, purmorphamine, db-cAMP, ascorbic acid, BDNF, and GDNF | >40Â days |
Takazawa et al. [37] | ESCs | ESC mediuma and then DMEM/F12, N2, NEAA, L-Glu, and heparin | Y27632, bFGF, and recombinant mouse Noggin | Embryoid bodies | Wnt3a-L-cell conditioned medium | RA, ascorbic acid, db-cAMP, and recombinant mouse Shh (C25II) | Neurobasal medium with N2, B27, Glu, and NEAA | Ascorbic acid, db-cAMP, RA, Shh, BDNF, GDNF, and IGF1 | >31Â days |
Wada et al. [46] | ESCs | ESC mediuma and FGF2 | Noggin and dorsomorphin | Neural rosette | DMEM/F12, N2B27 | Shh or SAG, RA | DMEM/F12, N2, B27, FGF2, and heparin | GDNF, BDNF, and NT3 | 38Â days |
FGF2, and EGF | |||||||||
Wichterle et al. [27] | mESCs | DFK5 medium (DMEM/F12, L-Glu, pen/strep, BME, and insulin-transferrin- selenium supplement | RA, Shh, Hedgehog agonist (Hh-Ag1.3), or hedgehog antibody (5E1) | Embryoid bodies | DMEM/F12 medium | - | DMEM/F12 | GDNF, BDNF, CNTF, and NT3 | ~25Â days |
Zeng et al. [47] | iPSC (ESCs as positive control) | hESC mediuma | N2 and heparin | Neural rosette | Neural mediumb | RA, Shh, and FGF8 | Neural basal medium, N2, and B27 | GDNF, BDNF, and IGF1 | >24Â days |