Table 1.
Components of the various media used in the selected differentiation protocols [85,86,87,88,89,90]. Many have similar base media, e.g., knockout (KO) DMEM/F12, as well as components routinely found in neuronal differentiation cultures such as N2 and B27. GlutaMax is a more stable form of L-glutamine. The addition and concentration of additional growth factors varies substantially between protocols.
| Medium 1 | Medium 2 | Medium 3 | Medium 4 | Medium 5 | Medium 6 |
|---|---|---|---|---|---|
| 80% KO DMEM/F12, 20% KSR, 1% NEAA 1 mM L-Glu 0.1 mM 2-ME |
KO DMEM/F12, 2% StemPro neural suppl., 20 ng/mL FGF-2, 20 ng/mL EGF |
N2 M., 10 ng/mL CNTF, 10 ng/mL FGF-2, 1 mM dBcAMP, 20 ng/mL NRG1 |
50% (Neurobasal M., MEM-NEAA, GlutaMAX, B27, 20 ng/mL FGF-2) + 50% (SDIA condition M, 10 μM Y-27632, 200 μM AA) |
MesenPRO M., 20 ng/mL NRG1 |
SDIA condition M., 10% KSR, 0.1 mM NEAA, 1 mM pyruvate, 0.1 mM 2-ME |
| Medium 7 | Medium 8 | Medium 9 | according to Kreitzer et al. [88] | ||
| KSR medium | N2 medium | ||||
| DMEM/F12, 20 ng/mL FGF-2, 1% N2, 2% B27, 0.05% BSA fraction V, 1% GlutMax, 1% MEM-NEAA, 110 µM 2-ME, 10 µM Y-27632 |
Advanced DMEM/F12 + Neurobasal M. (1:1 mix), 1% N2, 2% B27, 0.005% BSA, 2 mM GlutaMax, 0.11 mM 2-ME, 3 mM CT99021, 20 mM SB-431542 |
DMEM/low glucose, 1% FBS, 4 mM FK, 200 ng/mL NRG1, 100 nM all-trans RA, 10 ng/mL PDGF-BB |
Knockout DMEM, 15%KSR, 1% MEM-NEAA, 1% GlutaMax, 55 µM 2-ME |
DMEM/F12, 0.15% glucose, 1% N2, 20 µg/mL insulin, 5 mM HEPES |
|
AA: ascorbic acid. B27: B-27™ Supplement. BSA: bovine serum albumin. CNTF: ciliary neurotrophic factor. CT99021: 6-((2-((4-(2,4-Dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)pyrimidin-2-yl)amino)ethyl)amino)nicotinonitrile. dBcAMP: dibutyryl cyclic adenosine monophosphate. EGF: epidermal growth factor. FBS: fetal bovine serum. FGF-2: basic fibroblast growth factor. FK: forskolin. HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid. KSR: knockout serum replacement. L-Glu: L-glutamine. M: medium. N2: N-2 Supplement. NEAA: non-essential amino acid. NRG1: neuregulin 1. PDGF-BB: platelet-derived growth factor two B subunits. SB-431542: 4-[4-(1,3-benzodioxol-5-yl)-5-(2-pyridinyl)-1H-imidazol-2-yl]benzamide. SDIA: stromal cell-derived inducing activity. Suppl: supplement. 2-ME: 2-mercaptoethanol. Y-27632: (1R,4r)-4-((R)-1-aminoethyl)-N-(pyridin-4-yl)cyclohexanecarboxamide.