Method for inducing neuroblastoma cells to be differentiated to nerve cells
A neuroblastoma and neural stem cell technology, applied in the field of cell research, can solve the problems of long differentiation time and low neuron cell differentiation efficiency, and achieve the effect of inhibiting cell apoptosis
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[0024] (1) Preparation of neural stem cell conditioned medium
[0025] Human neural stem cells were purchased from Lonza Company and cultured in T-75 culture flasks with 15 ml of complete medium. Complete medium containing DMEM / F12 basal medium (Invitrogen), human recombinant epidermal growth factor (EGF, 20ng / mL) and basic fibroblast growth factor (bFGF, 20ng / mL) (R&D), B27 (serum replacement , 1:50, Invitrogen), heparin (5 μg / mL, Sigma), L-glutamine 2mM, and penicillin and streptomycin (1:100, Invitrogen). at 37°C with 5% CO 2 Incubate in a humidified incubator, and replace 50% of the medium with new complete medium every 3 days. When the neurospheres grow to a diameter of about 1 mm, they are cut and passaged under a microscope. When the culture medium is replaced each time, the old culture medium replaced is the neural stem cell conditioned medium. The conditioned medium was collected and filtered through a 0.22 μm pore size filter (Millipore), then centrifuged at 1000...
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