Method for culturing far-end pulmonary artery endothelial cell of rat

Abstract

The invention discloses a culture method of white rat distal pulmonary artery endothelial cells. The invention comprises the following steps: firstly, distal pulmonary arteries are obtained from a healthy white rat cardiopulmonary organization; secondly, the distal pulmonary artery endothelial cells are extracted from the distal pulmonary arteries, and the culture of primary cells is performed, to obtain the white rat distal pulmonary artery endothelial cells of the primary culture; thirdly, after the primary cells obtained in the step (2) is washed with PBS solution, 0.3 to 0.5 ml 0.1 to 0.2percent of trypsin solution is added to digest for 2 to 3 minutes at temperature 20 to 37 DEG C. when the cell retracts, and the cell interval is increased, the trypsin solution is resorped, 5 ml mixed culture solution II is added, the cells on the wall of a light blowing bottle is lightly blown, to ensure the cells to form cell suspension and to be inoculated into a new culture bottle, the once liquid change is performed every two days, until the cells grow to the logarithmic growth phase, the culture of continuous cells are finished. The method of the invention paves the condition for further deeply research the pulmonary circulation relevant disease and the small vessel disease and other pathogeny mechanism of the relevant disease, and provides the abundant material source.

Description

technical field [0001] The invention relates to a cell culture method, in particular to a culture method for rat distal pulmonary artery endothelial cells, belonging to the field of biotechnology. Background technique [0002] Pulmonary artery endothelial cells, especially distal pulmonary artery endothelial cells, are important cell materials for studying the pathogenesis of pulmonary circulation-related diseases, especially pulmonary hypertension and other diseases. Currently, the methods for culturing pulmonary artery endothelial cells include enzymatic digestion and tissue block patching. Most of the animal sources are large animals such as cattle and pigs. The cost is high, the quality of animals is difficult to control, and there are few sources. Moreover, the tissue patching method is not yet available for culturing cells. It has the disadvantages of long period and low success rate. However, rats are the most common, practical and convenient experimental animals in...

AI Technical Summary

Problems solved by technology

At present, the methods for culturing pulmonary artery endothelial cells include enzymatic digestion and tissue patch methods. The animal sources are mostly large animals such as cattle and pigs, which are expensive, difficult to control the quality of animals, and have fewer sources. It has the disadvantages of long cycle and low success rate

However, rats are the most common, practical and convenient experimental animals in the field of life science research, but there are few reports about the cell culture method using the distal pulmonary artery of rats as the source of endothelial cells. The reasons for the analysis mainly include the following reasons: 1. Rats are small in size and have smaller heart and lung tissues. It is too difficult and difficult to isolate the distal pulmonary artery of rats with the conventional method of isolating the distal pulmonary artery of large animals such as cattle and pigs: 2. The isolated distal pulmonary artery of the rat Too thin, it is not feasible to culture rat distal pulmonary artery endothelial cells with conventional large vessel enzyme perfusion digestion

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