A | B | |
DMEM F12 (w/o HEPES) /Neurobasal (1:1) | ||
N2 0.5% | 250 µL/50 mL | |
B27 1% | 500 µL/50 mL | |
NEAA 1% | 500 µL/50 mL | |
B-MercaptoEtOH 0.1% | ||
Heparin | 1 µg/mL (100 mg/mL) 1:100.000 | |
SB431542 | 10 µM (10 mM) 1:1000 | |
Noggin | 200 ng/mL (200 µg/mL) 1:1000 or (LDN 1:100.000) | |
CHIR99021 | 0.7 µM (3 mM) 1:4285 11.7 µL/50 mL | |
Rock Inhibitor (APOI) | 50 µM (10 mM) 1:200 |
A | B | |
DMEM F12 (w/o HEPES)/Neurobasal (1:1) | ||
N2 0.5% | 250 µL/50 mL | |
B27 1% | 500 µL/50 mL | |
NEAA 1% | 500 µL/50 mL | |
B-MercaptoEtOH 0.1% | ||
Heparin | 1 µg/mL (100 mg/mL) 1:100.000 | |
SB431542 | 10 µM (10 mM) 1:1000 | |
Noggin | 200 ng/mL (200 µg/mL) 1:1000 or (LDN 1:100.000) | |
CHIR99021 | 7.5 µM (3 mM) 1:400 | |
SHH-C25II | 100 ng/mL (100 µg/mL) 1:1000 | |
FGF8 | 100 ng/mL (100 µg/mL) 1:1000 | |
PMA | 1 µM (Stock: 1mM) |
A | B | |
Neurobasal | ||
N2 | 0.50% | |
B27 | 1% | |
Glutamax | 1% | |
NEAA | 1% | |
B-MercaptoEtOH | 0.1% | |
Insulin | 2.5 µg/mL (5 mg/mL) 1:2000 | |
Laminin | 200 ng/mL (100 µg/mL) 1:500 | |
SHH-C25II | 100 ng/mL (100 µg/mL) 1:1000 | |
CHIR99021 | 7.5 µM (3 mM) 1:400 | |
FGF8 | 100 ng/mL (100 µg/mL) 1:10.000 | |
PMA | 1 µM (Stock: 1 mM) |
A | B | |
Neurobasal | ||
N2 | (250 µL/50 mL) | |
B27 | (500 µL/50 mL) | |
Glutamax | 1% (500 µL/50 mL) | |
NEAA | 1% (500 µL/50 mL) | |
B-MercaptoEtOH | 0.1% (50 µL/50 mL) | |
BDNF | 10 ng/mL (20 µg/mL) 1:2000 | |
GDNF | 10 ng/mL (20 µg/mL) 1:2000 | |
Ascorbic Acid | 100 µM (200 mM) 1:2000 | |
db-cAMP | 125 µM (500 mM) 1:4000 | |
CHIR99021 | 7.5 µM (3 mM) 1:400 |
A | B | C | D | |
Company | Cat.Nr. | Name | Size | |
ThermoScientific | 12-587-010 | B27 | 10 ml | |
ThermoScientific | 17502-048 | N2 | 5 ml | |
R&D System | 3400-010-03 | Laminin I | 1 mg | |
Corning | 354230 | Matrigel | 10 ml | |
Selleckchem | S1049 | ApoI (Y-27632 2HCl) | ||
AppliChem | A0455,1000 | dbCAMP (C) | 1 mg | |
Sigma-Aldrich | 540220-5MG | Purmorphamine (PMA) | ||
R&D System | 1614 | SB 431542 | 10 mg | |
PeproTech | 450-02-500 | BDNF | 500 µg | |
PeproTech | 450-10-500 | GDNF | 500 µg | |
PeproTech | 100-25-500 | FGF8 | 500 µg | |
PeproTech | 100-45-100 | Sonic hedgehog | 100 µg | |
Sigma-Aldrich | A6964-100 ml | Accutase | 100 ml | |
Gibco | 11320074 | DMEM/F12 w/o HEPES | 500 ml | |
Gibco | 21103049 | Neurobasal Medium | 500 ml | |
Gibco | 11140035 | MEM NEAA | 100 ml | |
Gibco | 35050038 | Glutamaxx | 100 ml | |
Sigma | SML1046-5MG | CHIR 99021 | 3 mM | |
Sigma | 25556-4 | L-Ascorbic Acid | 200 mM | |
ThermoScientific | 12585014 | Insulin, human recomb. | 4 mg/ml | |
Axon | 1509 | LDN 193189HCl | 100 µM |