A method for inducing neural stem cells to differentiate into neurons in vitro
A neural stem cell and directional differentiation technology, applied in the field of inducing neural stem cells directional differentiation into neurons in vitro, can solve the problems of low neural stem cell differentiation ratio, low neuron purity, and no unified standard, and achieves the promotion of pure differentiation and high purity. , the effect of the simple method
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Embodiment 1
[0028] A method for inducing directional differentiation of neural stem cells into neurons in vitro, comprising the steps of:
[0029] (1) Primary culture of neural stem cells
[0030] Under sterile conditions, 14.5-day-pregnant ICR mice were sacrificed by dislocation of the cervical spine, soaked in 75% ethanol for 5-8 minutes, then transferred to an ultra-clean table for dissection, the fetal mice were taken out, and the brain cortex was separated, and then passed through 1×PBS After rinsing with DMEM / F12 solution, transfer to the growth medium containing 1% N2, 1% B27, 20ng / mL bFGF, 20ng / mL EGF, 97% DMEM / F12, and cut the brain tissue with dissecting scissors, Use a 1mL sterile pipette tip to blow repeatedly into a single-cell suspension. After the single-cell suspension is filtered through a 140-mesh nylon screen, transfer it to a 50ML centrifuge tube. The solution formula is: 1% N2, 1% B27, 20ng / mL bFGF, 20ng / mL EGF, 97% DMEM / F12) to adjust the cell density to 1×10 6 cel...
Embodiment 2
[0038] In step (3), the cell density of plating is 5×10 4 cells / mL, the rest are the same as in Example 1.
Embodiment 3
[0040] In step (3), the cell density of plating is 5×10 6 cells / mL, the rest are the same as in Example 1.
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