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Specification for hydrogen treatment of reperfusion injury of dermal microvascular endothelial cells

A technology of endothelial cells and microvessels, applied in the field of culture of dermal microvascular endothelial cells, can solve problems affecting the normal metabolism of cells

Inactive Publication Date: 2019-06-21
BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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Problems solved by technology

Because ischemia-reperfusion injury can lead to damage and even apoptosis of dermal microvascular endothelial cells, but the use of drugs or other methods has little effect and often affects the normal metabolism of cells. Ischemia-reperfusion injury, but does not affect the normal metabolism of cells, thereby reducing the reperfusion injury of dermal microvascular endothelial cells

Method used

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  • Specification for hydrogen treatment of reperfusion injury of dermal microvascular endothelial cells
  • Specification for hydrogen treatment of reperfusion injury of dermal microvascular endothelial cells
  • Specification for hydrogen treatment of reperfusion injury of dermal microvascular endothelial cells

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Embodiment 1

[0029] Dissolve hydrogen gas in serum-free ECM medium for 2 hours under 0.4MPa pressure to make it supersaturated. After decompression, hydrogen gas will be released. Use the same method to produce oxygen-saturated and carbon dioxide-saturated ECM medium. These three The medium is mixed in proportion (hydrogen medium: oxygen medium: carbon dioxide medium = 75%: 20%: 5%). During cell culture, put the mixed medium into the culture bottle and immediately use a hydrogen detector to detect hydrogen concentration, and then pass a well-proportioned mixed gas into the incubator, that is, hydrogen accounts for 75%, oxygen accounts for 20%, and carbon dioxide accounts for 5%, so that the concentrations of various gases in the medium are balanced and stable, and then the cells are added to the incubator. The culture bottle was closed and cultured for 24 hours. Under this condition, the hydrogen content of the medium was about 0.6MPa. Through this specification, hydrogen can be used in the...

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Abstract

The invention discloses a manufacture method of a hydrogen saturated medium, comprising dissolving hydrogen in a serum-free ECM (extracellular matrix) medium for 2 hours under the pressure of 0.4 MPa,saturating the medium, depressurizing to allow hydrogen to be released, manufacturing O2 and CO2 saturated ECM media in the same way, and mixing in a ratio of (H2:O2:CO2=75%:20%:5%). The invention also discloses a specification for treating reperfusion injury of dermal microvascular endothelial cells by hydrogen saturated medium and hydrogen; in cell culture, the mixed medium is placed in a culture bottle, hydrogen concentration is detected immediately with a hydrogen measurer; a mixed gas having 75% of hydrogen, 20% of oxygen and 5% of carbon dioxide is introduced into a culture box; the concentrations of gases in the medium are kept balanced and stable; cells are added in the culture bottle and cultured in enclosed manner for 24 hours; the hydrogen content of the medium under the condition is about 0.6 MPa.

Description

technical field [0001] The invention mainly relates to the use of hydrogen gas in the cultivation of dermal microvascular endothelial cells in cell experiments, especially for alleviating the ischemia-reperfusion injury of dermal microvascular endothelial cells. Background technique [0002] In 2013, "Molecular Hydrogen Biology" edited by Sun Xuejun was published by the Second Military Medical University, which explained in detail the impact of hydrogen on human health. He pointed out that hydrogen has three properties, one is effectiveness: it has anti-oxidation effect, and the other is Safety: In addition to the anesthetic effect lower than that of nitrogen, hydrogen has almost no adverse effects. The third is economical: the effective dose is not particularly high, and hydrogen has a strong penetrability, which can easily penetrate the cell membrane and reach the cytoplasm, Cell organelles such as mitochondria. [0003] Ischemia-reperfusion refers to the main factor that...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 林煌崔超李文志赵银花史艳宇曹建坤
Owner BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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