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Method for establishing hypoxia injury cell model

A technology of cell model and hypoxic injury, which is applied in the field of biomedicine to achieve the effect of low modeling cost and sensitive response to hypoxic stress

Inactive Publication Date: 2019-01-25
兰州军区兰州总医院
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a method for establishing a hypoxic injury cell model to solve the needs of the above-mentioned background technology. The advantages of being more sensitive, simple and stable can quickly simulate the hypoxic environment of cells and reflect the rapid development of cells in hypoxic environments. Peripheral oxygen partial pressure changes and the state of cell damage method issues

Method used

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  • Method for establishing hypoxia injury cell model
  • Method for establishing hypoxia injury cell model
  • Method for establishing hypoxia injury cell model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment one, refer to the attached Figure 2-3 .

[0038] Establishment of hypoxic injury model of rat liver cells (BRL)

[0039] 1. BRL cell culture conditions:

[0040] BRL cells were cultured in DMEM medium containing 10% fetal bovine serum (containing 1×105 IU / L penicillin and 1×105 IU / L streptomycin) in an incubator at 37°C and 5% CO2.

[0041] 2. Hypoxia treatment conditions for BRL cells:

[0042] BRL cells were cultured in DMEM high-glucose medium containing 10% fetal bovine serum. The hypoxic environment was 1% O2, 5% CO2, and 95% N2. The rest of the culture conditions were the same as normal cells. The hypoxic injury time was according to the experimental design requirements. conduct.

[0043] 3. Hypoxic injury experiment of BRL cells:

[0044] Three groups were designed in the experiment, namely control group, hypoxia treatment group 1 and hypoxia treatment group 2.

[0045] Digest BRL cells with 0.1% trypsin solution, make cell suspension, inoculate i...

Embodiment 2

[0054] Embodiment two, see appendix Figure 4-5 .

[0055] Determination of protein expression of hypoxia-inducible factor (HIF-1α) in rat liver cells (BRL)

[0056] 1. BRL cell culture conditions:

[0057] Described with specific embodiment 1.

[0058] 2. Hypoxia treatment conditions for BRL cells:

[0059] Described with specific embodiment 1.

[0060] 3. Hypoxic injury experiment of BRL cells:

[0061] Three groups were designed in the experiment, namely control group, hypoxia treatment group 1 and hypoxia treatment group 2.

[0062] Digest BRL cells with 0.1% trypsin solution, prepare cell suspension, inoculate in three six-well plates respectively, adjust the density to 1.0×106 / mL, add 3.0×105 cells per well, that is 33 microliters For the cell suspension, add 167 microliters of complete medium to 200 microliters, culture the cells for 24 hours, absorb the original medium after the cells are completely attached to the wall, and wash twice with PBS.

[0063] Control...

Embodiment 3

[0072] Embodiment three, see appendix Figure 6 .

[0073] Application of hypoxic injury model to screen and validate anti-hypoxic drugs

[0074] 1. BRL cell culture conditions:

[0075] Described with specific embodiment 1.

[0076] 2. Hypoxia treatment conditions for BRL cells:

[0077] Described with specific embodiment 1.

[0078] 3. BRL cell hypoxia model to verify the anti-hypoxic effect of acetazolamide:

[0079] Inoculate the BRL cell suspension in 96-well plates, adjust the density to 1.0×106 / mL, add 1.0×104 cells per well, that is, 10 microliters of cell suspension, add 90 microliters of complete medium to 100 microliters 1, culture the cells for 24 hours, after the cells are completely adhered to the wall, the original medium is sucked off, and the cells are washed twice with PBS.

[0080] Construct hypoxic injury cell model according to the method described in the present invention, promptly the cell adheres to the wall and grows on the 96-well plate, after w...

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Abstract

The invention discloses a method for establishing a hypoxia injury cell model in the field of biomedical technology. The method comprises the following steps: S1, cell culture; 2, cell plate; S3, medium replacement; S4, cell hypoxia culture: covering a layer of permeable membrane on the culture container and culturing in a modular culture chamber containing a certain gas component; S5, cell detection. The hypoxia injury cell model established by this method does not require the participation of experimental animals, and the modeling cost is low. It is more sensitive to hypoxia stress reaction,and can be used to explore hypoxia injury at the molecular biological level. It can provide conditions for the simulation of hypoxia experiment and the screening and verification of anti-hypoxia drugs.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for establishing a hypoxia-damaged cell model. Background technique [0002] Hypoxia refers to the pathological process of abnormal changes in the body's metabolism, function, and morphological structure due to insufficient oxygen supply or oxygen use disorders in tissues. The lack of oxygen in vital organs such as the brain and heart is also an important cause of death. In addition, insufficient oxygen supply to tissues due to a significant decrease in arterial blood oxygen content can lead to hypoxemia. [0003] The normal life activities of the body are inseparable from oxygen, and everything from cell metabolism to heart beating is closely related to oxygen. The body inhales oxygen, reaches the tissue through the blood, and is finally sensed and utilized by the cells. Therefore, the essence of hypoxia is a response and adaptive change of the cells to the hypoxic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00
CPCC12N5/00C12N2500/02
Inventor 王荣贾正平岳新瑞谢华李文斌鹿辉
Owner 兰州军区兰州总医院
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