Method for separating and cultivating endothelial cells of common carotid arteries of SD (Sprague Dawley) rats

Abstract

The invention discloses a method for separating and cultivating endothelial cells of common carotid arteries of SD (Sprague Dawley) rats. The method includes steps of 1), incising the common carotid arteries of the SD rats under sterile conditions; 2), cutting and segmenting the common carotid arteries, then stripping outer membranes from the common carotid arteries in PBS (poly butylenes succinate) liquor, longitudinally cutting the common carotid arteries, downwardly flatly spreading inner membranes in containers filled with digestive enzyme liquor and enabling the inner membranes of the common carotid arteries to be in sufficient contact with the digestive enzyme liquor in the containers; 3), vibrating the containers to allow the endothelial cells to fall off, removing the common carotid arteries after digestion is completed, adding fetal bovine serum into the containers and uniformly stirring the fetal bovine serum to obtain mixed liquid; 4), centrifuging the mixed liquid, removing supernatants to obtain the endothelial cells of the common carotid arteries, cultivating the endothelial cells of the common carotid arteries in endothelial cell cultivation media, transferring the endothelial cells to cultivation tanks to cultivate the endothelial cells after the endothelial cells reach log phases and then carrying out generation cultivation on the endothelial cells. The method has the advantages of high survival rate, little cell injury, economic efficiency, practicality, simplicity, feasibility and applicability to mass production in factories and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for separating and culturing SD rat common carotid artery endothelial cells. Background technique [0002] Vascular endothelial cells have a variety of physiological functions, can produce and secrete many biologically active substances, and play an important role in maintaining vasomotor and anticoagulation. The successful culture of vascular endothelial cells in vitro is the basis for studying the function of endothelial cells and their role in the occurrence and development of various diseases. Endothelial cells (EC) isolated from the common carotid artery of SD rats are widely used in experimental research. [0003] Endothelial cells have poor growth ability in vitro, and primary culture is not easy to succeed. In the past, mechanical separation and tissue block transplantation were mainly used. Cut the umbilical vein, scrape the endothelial cells with the back of a s...

AI Technical Summary

Problems solved by technology

Cut the umbilical vein, scrape the endothelial cells with the back of a scalpel, or use a separation tube with a nylon mesh, but it is difficult to control the strength of this method. If the force is too light, the number of cells obtained is very small; if the force is too heavy, it is easy to mix Other blood vessel wall cells such as fibroblasts, etc., and this method is easy to damage endothelial cells

In recent years, the separation of endothelial cells has been gradually replaced by enzymatic digestion. The cells obtained by this method are relatively pure and can better maintain their biological activity. However, trypsin has a strong ability to decompose the cell membrane and can destroy the integrity of the cells. , affect the viability of cells, and the endothelial cells obtained from digestion may contain a large number of fibroblasts, which will affect subsequent experiments. The collagenase digestion time is long, the cells are not fully dissociated and the damage is large, and the cells obtained are very few. Culture Poor cell viability

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