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Preparation method and application of acute lung injury cell model

A cell model and cell technology, applied in the field of medicine and biology, can solve problems such as doubts about objectivity, defects, and unclear characteristics of ATII cells

Active Publication Date: 2021-09-21
THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the same target, drugs rarely have dual regulatory activities, so this leads to doubts about the objectivity of using A549 as an ALI cell drug screening model
[0006] The reason may be that A549 has dual cell characteristics, and the ATII cell characteristics expressed by it are not obvious when used, which leads to obvious defects in its drug screening cell model for ALI, which easily leads to poor objectivity of evaluation results

Method used

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  • Preparation method and application of acute lung injury cell model
  • Preparation method and application of acute lung injury cell model

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1 A549 cell induction culture

[0023] experimental method:

[0024] A549 cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences at 37°C in 95% O 2 and 5% CO 2 The conditioned culture was subcultured once every two days, and the A549 cells in the logarithmic growth phase were taken, digested and collected, part of which was suspended with SABM medium (small airway basalmedium, Lonza), and the other part was suspended with RPMI 1640 medium (Hyclone , containing 10% volume of FBS) suspension (control group), and then adjust the concentration to 1×10 5 cells / ml spread in 6-well plates, 2ml per well, placed in an incubator and cultured for 1 day, the A549 cells cultured in SABM medium were randomly divided into groups, each group was added with different inducing compositions, and the control group was given the same amount of The RPMI1640 medium was cultured for 3 days before detection.

[0025] Detection Indicator:

[0026] 1) De...

Embodiment 2

[0041] Example 2 Preparation of A549 Cell Apoptosis Model Induced by Inflammatory Substances

[0042] According to the method of Example 1, Group 2, A549 cells were cultured, then the cells were digested and collected, and the cells were repeatedly centrifuged and washed 5 times with PBS to ensure that there was no induction medium residue, and then suspended with serum-free RPMI 1640 medium, and the concentration was 1 × 10 5 cells / ml spread in a 6-well plate, 2ml per well, for apoptosis and ROS detection, and then press 1×10 5 Cells / ml were spread in a 96-well plate, 100ul per well, for cell viability detection. Another uninduced A549 cells were also plated in 6-well plate and 96-well plate.

[0043] The induced A549 cells were divided into groups, with 5 holes in each group of 96-well plate and 3 holes in each group of 6-well plate; they were LPS group, PAF group, LPS+PAF group and induction control group respectively. The corresponding inflammatory inducers were added ...

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Abstract

The invention relates to a preparation method and application of a cell model of acute lung injury, belonging to the field of medicine and biology. It includes: 1) placing A549 cells in the induction medium for induction culture, then discarding the medium, digesting and harvesting the cells; 2) suspending the harvested A549 cells in the medium, placing them in a culture dish, and adding inflammatory substances Stimulation triggers an inflammatory response, and after 12-48 hours of continuous culture, it can be used for ALI drug screening.

Description

technical field [0001] The invention relates to a preparation method and application of a cell model of acute lung injury, belonging to the field of medicine and biology. Background technique [0002] Acute lung injury (Acute lung injury, ALI) is a common clinical emergency, characterized by progressively aggravated dyspnea and persistent hypoxemia, and its severe stage can develop into adult respiratory distress syndrome (Acuterespiratory distress syndrome, ARDS), ARDS is the main cause of acute respiratory failure in clinical critical illness, with a mortality rate as high as 50%. Common pathogenic factors mainly include: infection, inhalation of harmful substances, trauma, shock, poisoning, etc. The pathogenesis of ALI is relatively complex, and its pathological basis is alveolar capillary membrane damage caused by uncontrolled inflammatory response in the lung, which in turn leads to pulmonary edema and hyaline membrane formation. Alveolar type II epithelial cells are ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693C12N2500/24C12N2501/052C12N2501/10C12N2501/115C12N2501/999G01N33/5011G01N2500/10
Inventor 叶进燕卢瑶张丹李成业
Owner THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV