Coculture method of glial cells and neurons
A glial cell and neuron technology, applied in the field of cell culture and cell biology, can solve the problems of high cell culture cost, less material source, complicated process, etc., and achieve the effect of simplified steps, avoidance of use, and easy handling.
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[0037] This embodiment provides a method for co-cultivating glial cells and neurons, including the following:
[0038] (1) Reagent preparation:
[0039] 1. B27 (Invitrogen, Cat. No. 17504) is used for neuron culture
[0040] 2. Laminin (10 mg ml–1; e.g., Sigma, Cat. No. L2020), can be added to poly-lysine to coat the coverslip
[0041] 3. Gentamicin (10 mg ml–1; Invitrogen, Cat. No. 15710)
[0042] 4. Glutamax (200 mM; stable dipeptide of Ala-Gln; Invitrogen, catalog number 35050-061)
[0043] 5. NeurobasalA (Gibco catalog number 10888022), the main component of complete medium in contact co-culture
[0044] 6. Poly-D-Lys is used to coat sterile coverslips
[0045] 7.OptiPrep density 1.32 (Sigma, catalog number D1556) is used for density gradient centrifugation reagent preparation
[0046] 8. HABG preparation: culture medium containing 60ml HA, 1.2ml B27, 0.176ml GIn (final 0.5mM)
[0047] 9. NeurobasalA complete medium: its preparation requires 25ml NeurobasalA, 0.5ml B...
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