10 cm sterile culture plates |
Falcon |
353003 |
|
Non-Essential Amino Acids (NEAA) |
Thermofisher |
11140050 |
Working concentration 100x |
Supplement N – N2 Supplement |
Thermofisher |
17502048 |
Working concentration 100x |
L-Glutamine |
Thermofisher |
25030 |
Working concentration 100x |
Neurobasal |
Thermofisher |
21103049 |
Working concentration 1x |
Retinoic acid (RA) |
Sigma |
R2625 |
Dissolve 100 mg into 3.3 mL of DMSO to get 100 mM stock solution. Aliquot the stock 100 µL/tube and freeze at -80 ºC. Take 200 µL of 100 mM stock and dilute 10x (add 1.8 mL of DMSO) to make 10 mM stock. Aliquot 50 µL/tube and store at -80 ºC. Dilute at 1:10000 for use. (working concentration 2 µM). |
Polyornithine (PLO) |
Sigma-Aldrich |
P3655 |
Dissolve 100 mg in 1 mL of ddW to get 100 mg/mL stock solution. Aliquot the stock in 100 µL tubes. (working concentration 100 µg/mL) |
Laminin |
Thermofisher |
23017-015 |
Stock solution 1 mg/mL, working concentration 10 µg/mL (coating), 1 µg/mL (cell media) |
Ascorbic acid (ASAC) |
Sigma |
A4403 |
Dissolve 100 mg into 250 mL of dH2O to get 0.4mg/ml stock. Sterile filter through a 0.22 µm filter, aliquot and freeze at -80 ºC. Dilute at 1:1000 for use. (working concentration 0.4 µg/mL). |
Ciliary neurotrophic factor (CNTF) |
Peprotech |
450-13 |
Dissolve 100 µg in 1mL of sterile PBS, and then add 9 mL of sterile 0.1% BSA-PBS to 10 µg/mL. Aliquot and freeze at -80 ºC. Dilute at 1: 1000 for use. (working concentration 10 ng/mL). |
Compound E |
Abcam |
ab142164 |
Dissolve 250 µg in 2.0387 mL of DMSO to get a 250 µM stock. Aliquot and freeze at -80 ºC. Dilute 1:2000 for use. (working concentration 125 nM). |
DMEM/F12 |
Thermofisher |
113300 |
Working concentration 1x |
Glial cell line-derived neurotrophic (GDNF) |
Peprotech |
450-10 |
Dissolve 100 µg in 1mL of PBS to 100 µg/mL, and then add 9 mL of sterile 0.1% BSA-PBS to 10 µg/mL. Aliquot and freeze at -80 ºC. Dilute at 1: 1000 for use. (working concentration 10 ng/mL). |
ROCK-I nhibitor |
Peprotech |
1293823 |
Dissolve 5 mg in 1480 µL of dH2O to get 10 mM stock, aliquot and freeze at -80 ºC. Dilute at 1: 500 for use. (working concentration 20 µM). |
Pencillin/Streptomycin |
Thermofisher |
15140122 |
Working concentration 100x |
Purmorphamine (PMN) |
Millipore-Sigma |
540223 |
Dissolve 5 mg in 9.6 mL of DMSO to get 1 mM solution. Aliquot and freeze at -80 ºC. Dilute 1:1000 for use. (working concentration 1 µM). |
Recombinant human-brain-derived neurotrophic factor (BDNF) |
Peprotech |
450-02 |
Centrifuge briefly before reconstitution. Dissolve 100 µg in 1 mL of PBS to 100 µg/mL, and then add 9 mL of sterile 0.1% BSA-PBS to 10 µg/mL. Aliquot and freeze at -80 ºC. Dilute at 1: 1000 for use. (working concentration 10 ng/mL). |
Insulin-like growth factor 1 (IGF-1) |
R&D systems |
291-G1-200 |
Dissolve 200 µg in 2 mL of PBS to 100 µg/mL, then add 18 mL of 0.1%BSA-PBS to make 10 µg/mL stock and store at -80 ºC. Aliquot and freeze at -80 ºC. Dilute 10 µg/mL stock at 1: 1000 for use. (working concentration 10 ng/mL). |
Glial cell line-derived neurotrophic (GDNF) |
Peprotech |
450-10 |
Dissolve 100 µg in 1mL of PBS to 100 µg/mL, and then add 9 mL of sterile 0.1% BSA-PBS to 10 µg/mL. Aliquot and freeze at -80 ºC. Dilute at 1: 1000 for use. (working concentration 10 ng/mL). |
Retinoic acid (RA) |
Sigma |
R2625 |
Dissolve 100 mg into 3.3 mL of DMSO to get 100 mM stock solution. Aliquot the stock 100 µL/tube and freeze at -80 ºC. Take 200 µL of 100 mM stock and dilute 10x (add 1.8 mL of DMSO) to make 10 mM stock. Aliquot 50 µL/tube and store at -80 ºC. Dilute at 1:10000 for use. (working concentration 2 µM). |
Compound E |
Abcam |
ab142164 |
Dissolve 250 µg in 2.0387 mL of DMSO to get a 250 µM stock. Aliquot and freeze at -80 ºC. Dilute 1:2000 for use. (working concentration 125 nM). |
Supplement B – B27 Supplement |
Thermofisher |
21985023 |
Working concentration 50x |
Sterile cell culture hoods |
Baker Company |
SG-600 |
|
Table top cell culture centrifuge |
ThermoFisher |
75004261 |
Sorvall Legend X1R |
Waterbath |
ThermoFisher |
2332 |
Isotemp |
Humidity controlled Cell culture incubator |
ThermoFisher |
370 |
set to 37 ºC, 5 % CO2 |