Improved mixed enzyme digestion of cultured rat aortic smooth muscle cells.docVIP

 PAGE \* MERGEFORMAT 7 Improved mixed enzyme digestion of cultured rat aortic smooth muscle cells Authors: Licheng Wu, Li added, Zhu Li, Song Jian [Abstract] Objective To study the modified enzyme digestion of cultured rat aortic smooth muscle cells (VSMC) of the method for atherosclerosis and other vascular sclerosis disease pathogenesis and prevention research assistance. Extracted from rat thoracic aorta, using mixed-enzyme digestion, using with 10? S RPMI1640 culture medium. Observation of cell proliferation by cell count, inverted phase contrast microscope biological properties of vascular smooth muscle cells, immunohistochemistry for cell identification. The results, after digestion and muscle 24h adherent smooth muscle cells, the average adhesion rate of 76%, the first 5 d after the cell number increased rapidly to 11 d form a single layer, the light microscope, appear ‘Feng  Valley’-like growth; after passage , cell growth characteristics of no change, α  SMA immunohistochemical staining confirmed that cells are VSMC. Conclusion improved enzymatic digestion can be short, efficient and stable biological characteristics of cultured VSMC. [Keywords:] enzyme digestion method; smooth muscle cell; thoracic aorta; SD rats ABSTRACT: Objective To explore the cultivation of rat vascular smooth muscle cells (VSMC) by enzymatic dispersion which helps to provide ideals for the prevention and treatment of angiosclerotic diseases. Methods The thoracic artery was immediately removed and cut into fine tissue pieces, then was incubated in a mixed medium.After digestion, the cells were cultured in RPMI1640 with 20% FCS and then plated onto plastic tissue culture dishes.The proliferation of cell was assayed by cell counting, the feature of vascular smooth muscle cells was observed with inverted phase contrast microscope, and immunohistochemical method was used to indentify cells.Results After 24h of incubation, VSMCs began to stick to the wall of the incubation di