A method for isolating and culturing chicken embryonic pulmonary artery smooth muscle cells

Abstract

The invention relates to an isolated culture method for chicken embryo pulmonary arterial smooth muscle cells. The method comprises the following steps that a chicken embryo herringbone pulmonary blood vessel is isolated and kept wet through PBS, tiny connective tissue on the periphery of the vessel is trimmed and removed, the herringbone pulmonary blood vessel is divided into two vessels which are longitudinally cut at a time, a trypsin solution is used for digesting the longitudinally-cut vessels for 1-2 min at 15-37 DEG C, then a complete medium is used for stopping digestion to obtain a digested tissue block, the digested tissue block is cut into small tissue blocks with the size of 0.4-0.6 mm, and primary culture is carried out on the small tissue blocks in the complete medium. According to the isolated culture method for chicken embryo pulmonary arterial smooth muscle cells, operation is easy, the repeatability is good, the cultured cells are high in purity, the growth period is short, cells can climb out at the third day of culture, continuous cell culture can be carried out after 4 days of primary culture, and the cells grow vigorously.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for isolating and culturing chicken embryonic pulmonary artery smooth muscle cells. Background technique [0002] In vitro experiments using primary cells can directly study the responses of primary cultured cells to stress factors such as growth factors, lipoprotein oxidation, and matrix protein synthesis under specific conditions, without being affected by other factors (such as blood pressure). Chicken pulmonary artery smooth muscle cells are important materials for studying the pathogenesis of diseases related to chicken cardiopulmonary health, especially broiler ascites syndrome (also known as pulmonary hypertension syndrome). Primary cultured pulmonary artery smooth muscle cells are capable of conducting research on signal transduction, protein phosphorylation, cell proliferation and hypertrophy, and are an important model for studying chicken cardiovascular health and...

AI Technical Summary

Problems solved by technology

If only the pulmonary aorta is used as the experimental material, a large amount of chicken embryos will be consumed

These two methods can be used to separate and culture low-day-old chicken pulmonary artery smooth muscle cells, but when these two methods are applied to the culture of chicken embryonic pulmonary artery smooth muscle cells, the results of the culture often cannot meet the requirements, resulting in poor culture efficiency and low purity The reasons may include the following points: 1. The body of the chicken embryo is small, and the lung tissue is small, embedded in the sternum and ribs, and is in a relatively closed space. It is difficult to separate and obtain a complete lung for enzyme digestion and culture; 2. The isolated The pulmonary artery of the chicken embryo is short and thin (<1cm), and the fibrous connective tissue is adhered around the blood vessel. It is difficult to separate the media for patch culture; 3. It is difficult to guarantee the purity of the primary cultured chicken pulmonary artery smooth muscle cells by enzymatic digestion. Adherent culture can improve cell purity, but there are many steps and long growth cycle

Although the sticking method (cutting the pulmonary aorta into pieces, sticking the pieces in a culture dish and adding culture medium) is simple to operate, but when the primary chicken embryonic pulmonary artery smooth muscle cells are cultured, the pulmonary arteries are slender and thin, and the tissue pieces are easy to stick into groups. Risk of fibroblast contamination

Images