Intracranial and spinal excitotoxicity injured cell network model and building method and application thereof

A cell network, exciting technology, applied in the field of biomedicine, which can solve the problems of insufficiency, loss of cell line characteristics, low degree of tight junction, etc.

Inactive Publication Date: 2015-05-20
DALIAN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although cell lines have the advantages of stable characteristics and easy acquisition, cell lines often lose a lot of original characteristics, and the impact on experimental results is difficult to estimate
In the second type of model, some cells are induced and differentiated from their own precursor cells. Although some of th...

Method used

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  • Intracranial and spinal excitotoxicity injured cell network model and building method and application thereof
  • Intracranial and spinal excitotoxicity injured cell network model and building method and application thereof
  • Intracranial and spinal excitotoxicity injured cell network model and building method and application thereof

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Embodiment 1

[0029] 1. Preparation of main reagents

[0030] 1. N complete culture medium:

[0031] N basal culture medium (Gibco 10888022) 95% (V / V), B 27 (Invitrogen 7504044) 2% (V / V), penicillin streptomycin 1% (V / V) and L-glutamine 2mM were mixed according to the above ratio, and placed in 37°C, 5% CO before use 2 Incubate for more than 30 minutes in the cell culture incubator.

[0032] 2. N planting medium:

[0033] DMEM high-glucose medium (Gibco 11960044) 90% (V / V), fetal bovine serum 10% (V / V), penicillin streptomycin 1% (V / V) and L-glutamine 2mM according to the above Proportion to mix, put in 37°C, 5% CO before use 2 Incubate for more than 30 minutes in the cell culture incubator.

[0034] 3. N culture medium:

[0035] N basal culture medium (Gibco 0050128DJ) 95% (V / V), B 27 2% (V / V), penicillin 1% (V / V) and L-glutamine 2mM were mixed according to the above ratio.

[0036] 4. N glutamic acid-glycine joint exposure culture medium:

[0037] N basal culture medium (Gibco 00...

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Abstract

The invention belongs to the technical field of biological medicines and particularly relates to an intracranial and spinal excitotoxic injury cell network model and a building method and application thereof. The cell network model comprises primary homologous central nervous system capillary endothelial cells, astrocytes and nerve cells, stereoscopic coculture is carried out according to in vivo relative space position and quantity relationship of the primary homologous central nervous system capillary endothelial cells, the astrocytes and the nerve cells, and the environment of the central nervous system in a body in the case of intracranial and spinal excitotoxic injury is simulated by using a common cell culture system and glutamic acid-glycine cell culture fluid/normal cell culture fluid. The invention puts forwards a pharmaceutical research mode of drug administration through plug-in central nervous system capillary endothelial cell culture liquid for the first time, is different from a conventional in vitro pharmaceutical research mode in which drug directly acts on a target cell(s) in the central nervous system and is more according with the pharmacokinetics of a living body. Therefore, the intracranial and spinal excitotoxic injury cell network model and the building method thereof can be used for deep research on generation and development of excitotoxic injury; and moreover, a new cell network model can be provided for clinical diagnosis markers and medicine research and evaluation.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a primary neuron formed by central nervous system capillary endothelial cells (endothelial cells, E), astrocytes (astrocytes, A) and neurons (neurons, N). Brain and spinal cord excitotoxic injury cell network model based on homologous three-cell four-dimensional model and its construction method and application. Background technique [0002] Among the central nervous system diseases, intractable epilepsy, acute ischemic stroke, amyotrophic lateral sclerosis and Alzheimer's disease caused by excitotoxicity have seriously endangered human life and health. And this kind of brain and spinal cord excitotoxicity-related diseases have attracted more and more attention because of their complexity and refractory. Although a large number of researchers are currently working on the research and development of drugs for this type of disease, the effects of the above-mentione...

Claims

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Application Information

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IPC IPC(8): C12N5/0793C12N5/079C12Q1/02
Inventor 孙长凯郑铁铮杨晓寒赵慧彭金咏齐艳许丽娜范明孟醒李彧媛
Owner DALIAN MEDICAL UNIVERSITY
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