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In-vitro foam cell formation model and application thereof in asymptomatic carotid atherosclerosis study

A foam cell and model technology, applied in the field of biotechnology and medical research, can solve problems such as large individual differences, complex mechanisms and processes, and long time.

Inactive Publication Date: 2018-01-26
UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above methods take a long time, and the atherosclerosis model is constructed at the animal level, the individual differences are large, and the mechanism and process of formation are relatively complicated

Method used

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  • In-vitro foam cell formation model and application thereof in asymptomatic carotid atherosclerosis study
  • In-vitro foam cell formation model and application thereof in asymptomatic carotid atherosclerosis study
  • In-vitro foam cell formation model and application thereof in asymptomatic carotid atherosclerosis study

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1. Under the conditions of different tensile strength and different concentration of low-density lipoprotein, the formation of foam cells is induced

[0073] The chip device used in the present invention is a chip device independently developed and designed by the inventor (ie, a mechanically stretchable microfluidic chip cell culture device in the patent number ZL201310043685.3, hereinafter referred to as a cell culture device) to prepare foam cells. After cleaning and drying the cell culture device, use an autoclave for sterilization. After drying again, immediately transfer to the ultra-clean workbench for use. Second, place the cell culture device in a sterile cell culture dish (10 cm in diameter) before inoculating the cells, and use fibronectin to coat the bottom membrane of the cell growth chamber in the cell culture device to increase blood vessels Smooth muscle cell adhesion ability. After the cells adhere to the wall, adding a medium containing 50μg / ml as...

Embodiment 2

[0115] Example 2. Atmospheric inhalable particulate matter (PM 2.5 ) Influence on the formation of foam cells in pre-atherosclerosis

[0116] Epidemiological investigations show that PM 2.5 Related to the morbidity and mortality of cardiovascular disease. Studies in the population found that the thickness of the carotid artery-media thickness of the people living in the arteries 2.5 Every 23μg / m increase 3 , The thickness of the carotid artery intima-media increased twice as much as that of the control group, which shows that people are exposed to PM 2.5 It is closely related to atherosclerosis.

[0117] In this example, PM2.5 is taken as an example to illustrate that the foam cells provided by the present invention form an in vitro model for investigating PM 2.5 Influence on the formation of foam cells in pre-atherosclerosis.

[0118] First, collect the PM of Beijing in winter 2.5 . PM that will be collected on filter paper 2.5 Weigh, dissolve in ultrapure water, make a 5mg / ml solu...

Embodiment 3

[0121] Example 3. The effect of atorvastatin on foam cell formation in a multi-cell co-culture system

[0122] Currently, the most commonly used drugs in the treatment of atherosclerotic diseases are statins, of which atorvastatin is the most commonly used. A large number of research reports have been carried out on the lipid-lowering mechanism of statins, which have been clinically verified. With the deepening of research, it has been discovered that statins have the functions of improving endothelial cells, anti-inflammatory, inhibiting smooth muscle cell proliferation, anti-thrombosis and anti-platelet in addition to their lipid-lowering effects. The early sign of atherosclerosis is endothelial cell dysfunction, and the increase of low-density lipoprotein concentration is an important factor in inducing endothelial cell dysfunction. The inventor of the present invention added low-density lipoprotein to the multi-cell co-culture system. (25μg / ml), aggravate endothelial cell dy...

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Abstract

The invention discloses an in-vitro foam cell formation model and application thereof in asymptomatic carotid atherosclerosis study. A construction method of the foam cell formation model provided bythe invention comprises the following steps: sequentially culturing, from bottom to top, smooth muscle cells, endothelial cells and mononuclear cells in a cell culture device capable of performing mechanical stretching so as to obtain a multi-cell co-culture system; treating the multi-cell co-culture system by utilizing a low density lipoprotein, applying a tension effect to induce formation of foam cells by virtue of the cell culture device capable of performing mechanical stretching, thus obtaining the foam cell formation model during early occurrence of asymptomatic carotid atherosclerosis.According to the method disclosed by the invention, consumption of an animal model can be reduced, expenditure is saved, an economic and high-efficiency in-vitro screening model is provided for carotid atherosclerosis resistant new drug screening, and the in-vitro foam cell formation model has the advantages of being simple in preparation method, convenient in experimental operation, low in timeconsumption and stable in performance.

Description

Technical field [0001] The invention relates to an in vitro foam cell formation model in the field of biotechnology and medical research and its application in atherosclerosis research. Background technique [0002] The blockage of blood vessels caused by atherosclerosis can lead to various cardiovascular diseases, including myocardial infarction, stroke, cerebral hemorrhage, and sudden death. The World Health Organization's statistical report shows that the death rate due to cardiovascular disease ranks first in the world, and this trend is increasing. As the incidence of atherosclerosis continues to rise and become younger, the resulting cardiovascular disease has brought a heavy burden to families and society, and has become a public health issue of general concern worldwide. Atherosclerosis is a chronically developing disease characterized by lipid deposition, inflammatory cell infiltration and fibrous tissue hyperplasia in large arteries. Atherosclerosis generally progress...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/077C12N5/0786C12M1/12C12M1/34C12M1/42C12M3/06C12Q1/02
Inventor 丁永胜顾晓阳
Owner UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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