Method for culturing rat airway smooth muscle cells

Abstract

The invention relates to a method for culturing rat airway smooth muscle cells. The method is characterized in that: the method includes two steps, i.e. obtaining primary-cultured rat airway smooth muscle cells and obtaining subcultured rat airway smooth muscle cells, wherein the step 1 digests the rate airway media with digestive solution under the temperature of 30 DEG C to 45 DEG C for 14 to 30 minutes, and then collects and primarily cultures the airway smooth muscle cells; and the digestive solution is prepared from HBSS balanced salt solution, type I collagenase, bovine serum albumin and dithiothreitol according to proportion. The culturing method has the advantages of stable technique, good repeatability, high cultured cell purity, uniform forms and well growth; and the digestive solution with the unique formula controls the digestion time of the airway smooth muscle layer at 14 to 30 minutes, thus overcoming the following defects that: in the prior art, since the rate media smooth muscle layer is thinner than the aorta and the smooth muscle cells are less than the cells of the aorta and the pulmonary artery, the eruption time of the airway smooth muscle cells is long, and growth is slow.

Description

technical field [0001] The invention relates to a cell culture method, in particular to a method for culturing rat airway smooth muscle cells, and belongs to the field of biotechnology. Background technique [0002] Asthma and other respiratory diseases are characterized by airway hyperresponsiveness and reversible airway obstruction, and abnormal airway changes are important pathological features of asthma. Airway smooth muscle cells are an important cell material for exploring the pathogenesis of respiratory diseases such as COPD and asthma. The cultivation of airway smooth muscle cells in vitro and the establishment of cell lines are the basis for the study of cell biological behavior and the study of disease pathogenesis and prevention methods . Therefore, it is of great significance to find a simple and efficient smooth muscle cell culture method for the research of respiratory diseases. At present, the methods for culturing airway smooth muscle cells mainly include p...

AI Technical Summary

Problems solved by technology

Among them, although the patch method is simple to operate, the cycle is too long, and the separation requirements for tissues are high. When the separation is not good, it is difficult for cells to climb out.

Moreover, the cell culture using rat airway as the source of smooth muscle cells is limited due to the following reasons: 1. Rats are smaller in size and have smaller heart and lung tissues. 2. The isolated rat trachea is less elastic than the arteries, with more fibrous connective tissue around it and tight adhesion, and there is no obvious excessive boundary between the adventitia, media and intima; 3. The smooth muscle layer of the media is relatively The aorta is thin, and there are fewer smooth muscle cells than the aortopulmonary artery. The general method of culturing airway smooth muscle cells takes a long time to erupt and grows very slowly.

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