动物造模 z-bio 2022-07-19 351

　　Objective: This paper reports an improved method for the isolation of rat thoracic aortic vascular smooth muscle cells based on compound enzymatic digestion.

　　Methods: The thoracic aorta was preliminarily digested with collagenase and trypsinase to remove the inner and outer membranes, and then the medial membrane was subjected to secondary digestion to release vascular smooth muscle cells.

　　RESULTS: The cells grew adherent to the wall after isolation, and most of them had a spindle-shaped shape, showing a typical "peak-valley" growth state, and could be passaged within 1 week; the molecular marker genes of various contractile vascular smooth muscle cells were all found in the isolated and cultured cells. Moderately high expression; more than 95% of cells were significantly positive for alpha smooth muscle actin (α-SMA) and myosin-II.

　　Conclusion: This separation method is simple, easy to implement, and has good repeatability. The obtained cells have good viability and high purity, and can ensure a large number of contractile vascular smooth muscle cells in a short time.