Acute brain and spinal ischemia/reperfusion injury cellular network model as well as construction method and application thereof

A technology of reperfusion injury and cell network, applied in the field of cell network model and construction of acute ischemia/reperfusion injury of the brain and spinal cord, which can solve the problems of low degree of tight connection and unsatisfactory

Inactive Publication Date: 2015-01-28
DALIAN MEDICAL UNIVERSITY
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  • Abstract
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  • Claims
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Problems solved by technology

However, the tight junction degree of E obtained according to the construction method of this model is often low, which cannot meet the requirements of injury experiments and drug research.

Method used

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  • Acute brain and spinal ischemia/reperfusion injury cellular network model as well as construction method and application thereof
  • Acute brain and spinal ischemia/reperfusion injury cellular network model as well as construction method and application thereof
  • Acute brain and spinal ischemia/reperfusion injury cellular network model as well as construction method and application thereof

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Embodiment 1

[0030] 1. Preparation of main reagents

[0031] 1.N complete culture medium:

[0032] Put N basic culture medium (Gibco 10888022) 95% (V / V), B 27 (Invitrogen 7504044) 2% (V / V), penicillin 1% (V / V) and L-glutamine 2mM are mixed according to the above ratio, put in 37℃, 5% CO before use 2 Incubate in a cell incubator for more than 30 minutes.

[0033] 2.N planting culture solution:

[0034] Combine DMEM high glucose culture medium (Gibco 11960044) 90% (V / V), fetal calf serum 10% (V / V), penicillin 1% (V / V) and L-glutamine 2mM according to the above Proportion to mix, put in 37℃, 5% CO before use 2 Incubate in a cell incubator for more than 30 minutes.

[0035] 3.N sugar-free culture medium:

[0036] Put N sugar-free basic culture medium (Gibco 0050128DJ) 95% (V / V), B 27 2%(V / V), penicillin 1%(V / V) and L-glutamine 2mM are mixed according to the above ratio, put in 37℃, 95%N before use 2 , 5% CO 2 Incubate for more than 12 hours in the cell culture box of the hypoxia control system.

[0037] ...

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Abstract

The invention belongs to the technical field of biological medicines and in particular relates to an acute brain and spinal ischemia/reperfusion injury cellular network model as well as a construction method and application thereof. The cellular network model comprises primary isogenous central nervous system capillary endothelial cells, astrocytes and neurons, wherein the endothelial cells, astrocytes and neurons are three-dimensionally co-cultured according to the relative spatial position and quantity relations of the endothelial cells, astrocytes and neurons in vivo; the environment of oxygen and glucose deprivation/reoxygenation and recovery of glucose in a central nervous system during acute brain/spinal ischemia/reperfusion injury in vivo is simulated by utilizing a low oxygen regulation system/ordinary cell culture system and glucose-free cell culture fluid/normal cell culture fluid. The cellular network model has the beneficial effects that a medicine research mode of adopting the central nervous system capillary endothelial cell culture fluid in lumens of inserts for administration is put forward for the first time, so that the cellular network model further conforms to the organism pharmacokinetics; the cellular network model is used for further research of acute brain/spinal ischemia/reperfusion injury emergence and development mechanisms; the new cellular network model is provided for research and evaluation of clinical diagnostic markers and medicines.

Description

Technical field [0001] The present invention belongs to the field of biomedicine technology, and particularly relates to a primary generation formed by central nervous system capillary endothelial cells (E), astrocytes (astrocytes, A) and neurons (neurons, N) A cell network model of brain and spinal cord acute ischemia / reperfusion injury based on a homologous three-cell four-dimensional model and its construction method and application. Background technique [0002] Acute ischemic stroke of the brain and spinal cord has the characteristics of high morbidity, high mortality, high disability and high recurrence rate. It is a major medical problem that seriously endangers human life and health. Clinically, acute drug delivery after the onset of the disease and preventive drug delivery to high-risk groups have been used to intervene in acute ischemic stroke of the brain and spinal cord, but with little effect. [0003] In recent years, there have been more and more researches on acute...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/079C12N5/0793C12Q1/02
Inventor 孙长凯范明杨晓寒吴丽颖郑铁铮刘淑红蔡楠赵永岐洪浩魏盼周孝峰
Owner DALIAN MEDICAL UNIVERSITY
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