Nerve stem cell special-purpose culture medium and culture method thereof
A technology of neural stem cells and culture methods, applied in the direction of nervous system cells, animal cells, vertebrate cells, etc., can solve the problems of low survival rate of neural stem cells, complex serum components, unstable quality, etc., to promote adhesion and proliferation, Maintain high activity rate, reasonable and effective formula
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Embodiment 1
[0029] A special medium for neural stem cells, the medium is composed of DMEM / F12 culture fluid and solute, the solute and its concentration in DMEM / F12 culture fluid are: 300 μg / ml 2′,3′-di Deoxyadenosine-5-triphosphate, 5 μg / ml N2 Additives, 15ng / ml stem cell growth factor, 5mg / ml thioglycerol, 50ng / ml nerve growth factor, 0.05mmol / ml acetylcholine, 10mg / ml carboxymethylcellulose sodium, 0.005mmol / ml retinoic acid and 0.110mmol / ml folic acid.
Embodiment 2
[0031] A special medium for neural stem cells, said medium is composed of DMEM / F12 culture fluid and solute, said solute and its concentration in DMEM / F12 culture fluid are: 350μg / ml 2′,3′-di Deoxyadenosine-5-triphosphate, 8 μg / ml N 2 Additives, 22ng / ml stem cell growth factor, 6mg / ml thioglycerol, 63ng / ml nerve growth factor, 0.06mmol / ml acetylcholine, 18mg / ml sodium carboxymethylcellulose and 0.15mmol / ml folic acid.
Embodiment 3
[0033] A method for culturing neural stem cells, comprising the steps of:
[0034] 1) The isolated brain nerve tissue is obtained under sterile conditions, and rinsed with PBS buffer containing 5% double antibody;
[0035] 2) The brain nerve tissue obtained in step 1) is cut into 1mmol 3 Fragments, add DMEM / F12 medium containing 0.5mg / ml type I collagenase, 0.5mg / ml type IV collagenase and 0.1mg / ml deoxyribonucleic acid I, at 8 ℃ CO 2 Digest and decompose in the incubator for 20 hours to obtain tissue suspension;
[0036] 3) Centrifuge the tissue suspension obtained in step 2), remove the supernatant, add DMEM / F12 medium to wash 3 times, add cell digestion solution, digest at 35°C for 57min, gently pipette and mix to stop the digestion, and place in DMEM Rinse in / F12, pass through a 200-400 mesh cell sieve, collect the filtrate, centrifuge and discard the supernatant to obtain a single cell suspension;
[0037] 4) Add the special medium for neural stem cells to the single ...
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