Vendor | Cat# | vol | ||||
phenol-free DMEM/F12 (1:1) | Thermo Fisher | 11039021 | 92.5 | mL | ||
insulin (0.02 mg/ml) | ITS-G (100X stock) | Thermo Fisher | 41400045 | 1 | mL | |
holo-transferrin (0.011 mg/ml) | ||||||
sodium selenite (13.4 ug/ml) | ||||||
B27 (2% v/v) (50X stock) | Thermo Fisher | 17504044 | 4 | mL | ||
N2 (0.5%, v/v) (100X stock) | Thermo Fisher | 17502048 | 0.5 | mL | ||
monothioglycerol (200 uM) | 11.5 M | Sigma Aldrich | M1753-100mL | 1.75 | uL | |
Glutamax (1X) (100X stock) | 100X | Thermo Fisher | 35050061 | 1 | mL | |
non-essential amino acids (NEAA; 1X) (100X stock) | 100X | Thermo Fisher | 11140050 | 1 | mL | |
Pen/ Strep | 100X | Thermo Fisher | 15140-122 | 1 | mL | |
(additional insulin (5 ug/mL)) we do not add it | Sigma Aldrich | I9278-5mL | 47 | uL |
Vendor | Cat# | dilution | ||||
iMGL diff base medium | ||||||
IL-34 (100 ng/mL) | 500 ug/mL in H2O | Peprotech | 200-34 | 1:5000 | ||
TGFb-1 (50 ng/mL) | 100 ug/mL in 10mM Citric Acid | Peprotech | 100-21 | 1:2000 | ||
M-CSF (25 ng/mL) | 100 ug/mL in H2O | Peprotech | 300-25 | 1:4000 |
Vendor | Cat# | dilution | ||||
iMGL Complete medium | ||||||
CD200 (100 ng/mL) | 100 ug/mL | Novoprotein | C311-50ug | 1:1000 | ||
CX3CL1 (100 ng/mL) | 100 ug/mL | Peprotech | 300-31 | 1:1000 |
iPSC Cell Culture – Maintenance and Expansion
A | B | C | D | E | F | |
Component | Stock Concentration | Final Concentration | Vendor | Catalog # | Volume | |
Phenol-free DMEM/F12 (1:1) | Thermofisher | 11039021 | 462.4 mL | |||
Insulin (0.02 mg/mL), Holo-transferrin (0.011 mg/mL), Sodium selenite (13.4 ug/mL) (ITS-G Solution) | 100X | 2X | Thermofisher | 41400045 | 10 mL | |
B27 | 50X | 2% V/V | Thermofisher | 17504044 | 10 mL | |
N2 | 100X | 0.5% V/V | Thermofisher | 17502048 | 2.5 mL | |
Monothioglycerol | 11.5M | 400 uM | Sigma Aldrich | M1753-100ML | 17.4 uL | |
Non-Essential Amino Acids (NEAA) | 100X | 1X | Thermofisher | 11140050 | 5 mL | |
Glutamax | 100X | 1X | Thermofisher | 35050061 | 5 mL | |
Pen/Strep | 100X | 1X | Thermofisher | 15140-122 | 5 mL | |
Recombinant Human Insulin | 20 mg/mL | 5 ug/mL | Sigma Aldrich | I2643 | 125 uL |
A | B | C | D | E | |
Vendor | Cetalog # | Dilution | |||
iMGL Diff Base Medium | |||||
IL-34 (100 ng/ml) | 500 ug/mL in H20 | Peprotech | 200-34 | 1:5000 | |
TGFb-1 (50 ng/ml) | 100 ug/mL in 10mM Citric Acid | Peprotech | 100-21 | 1:2000 | |
M-CSF (25 ng/ml) | 100 ug/mL in H20 | Peprotech | 300-25 | 1:4000 |
A | B | C | D | E | |
Vendor | Catalog # | Dilution | |||
iMGL Complete Medium | |||||
CD200 (100 ng/mL) | 100 ug/mL | Novoprotein | C311-50ug | 1:1000 | |
CX3CL1 (100 ng/mL) | 100 ug/mL | Peprotech | 300-31 | 1:1000 |