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.

Pending Publication Date: 2022-04-29
GUIZHOU MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The existing contact co-cultivation often needs to extract the two kinds of cells separately, culture them separately and then combine them for co-cultivation. The cost of cultivation is high, and the yield is relatively low

Method used

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  • Coculture method of glial cells and neurons
  • Coculture method of glial cells and neurons

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Embodiment 1

[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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Abstract

The invention discloses a glial cell and neuron co-culture method, relates to the technical field of cell culture, and solves the technical problems of few sources of existing contact type co-culture materials, high culture cost, complex culture process and low survival rate. The method mainly comprises the following steps: collecting glial cells and neurons from brain tissues of adult rats or elderly rats, carrying out combined inoculation, not changing the liquid in the early stage of inoculation, sucking half of suspension cells and a culture medium into another hole when clear suspension cell balls are observed, complementing with a complete culture medium, continuing to culture, continuously distributing the suspension cells and the culture medium, and continuously culturing the cells and the culture medium. The cells are cultured for 25-30 days until the cell density meets the contact type co-culture requirement, and the glial cells and the neurons are subjected to contact type co-culture after being cultured for 25-30 days, so that a neural network and a glial background are relatively stably formed; adult rats or old rats are taken as sources, the sources are easy, the cost is low, the repeatability is good, cells with a certain yield can be obtained, the survival time of the cells is long, and the method is suitable for long-time observation.

Description

technical field [0001] The present invention relates to the technical field of cell biology, more specifically to the technical field of cell culture. Background technique [0002] The existing contact co-cultivation often needs to extract the two kinds of cells separately, culture them separately and then combine them for co-cultivation. The cultivation cost is high and the yield is relatively low. Contents of the invention [0003] The object of the present invention is to provide a method for co-cultivating glial cells and neurons in order to solve the above technical problems. [0004] The present invention specifically adopts the following technical solutions in order to achieve the above object: [0005] A method for co-cultivating glial cells and neurons, comprising the following steps: [0006] S1: Extract neurons and glial cells from the brain tissue of adult mice or aged mice, and obtain the second layer of cells and the third layer of cells by gradient densit...

Claims

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Application Information

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IPC IPC(8): C12N5/079C12N5/0793
CPCC12N5/0619C12N5/0622C12N2502/081C12N2502/086C12N2509/00C12N2509/10
Inventor 刘艳洁傅晓丽
Owner GUIZHOU MEDICAL UNIV
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