Table 1 Details of different cell culture media used
Media type | Media composition |
|---|---|
Skeletal muscle growth media | SKGM2 medium (Lonza) + bFGF (10 ng/mL, Thermo Fisher, PHG0023) |
Skeletal muscle differentiation media | DMEM/F12 (Thermo Fisher, 11,330), ITS-G (Thermo Fisher, 41,400,045), N2 supplement (Thermo Fisher, 17,502,048), Pen/Strep, and L-glutamine supplemented with 10-µM TGF-b inhibitor SB-431542 (Tocris, 1614) and 10 ng/ml IGF-1 (R&D Systems, 291-G1) |
Coculture media or motor neuron media | Neurobasal + 37‐ng/ml hydrocortisone, 2.2‐μg/ml isobutylmethylxanthine, 10‐ng/ml brain‐derived neurotrophic factor, 10‐ng/ml ciliary neurotrophic factor, 10‐ng/ml CT‐1, 10‐ng/ml glial cell‐derived neurotrophic factor, 2% B‐27, 20‐ng/ml nerve growth factor, 20‐μM mitotic inhibitors, 2‐mM L‐glutamine, 417‐ng/ml forskolin, 1‐mM sodium pyruvate, 0.1‐mM β‐mercaptoethanol, and 2.5‐g/L glucose |
Vascular media | Vascular cell basal medium (ATCC PCS-100–030) + endothelial growth kit-VEGF (ATCC PCS-100–041) comprising of the following: rh VEGF: 5 ng/mL, rh EGF: 5 ng/mL, rh FGF basic: 5 ng/mL, rh IGF-1: 15 ng/mL, L-glutamine: 10 mM, heparin sulfate: 0.75 units/mL, hydrocortisone: 1 µg/mL, ascorbic acid: 50 µg/mL, fetal bovine serum: 2% |
Triculture media | Skeletal muscle differentiation tool (ATCC PCS-950–050) + coculture media + vascular media = 1:1:1 |