Preparation of primary cultured mesenteric artery smooth muscle cells for fluorescent imaging and physiological studies
Preparation of primary cultured mesenteric artery smooth muscle cells for fluorescent imaging and physiological studies
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DOI：
10.1038/nprot.2006.425
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Blaustein, Mordecai P.
中科院分区：
文献类型：
--
作者：
Golovina, Vera A.;Blaustein, Mordecai P.
在本方案中，我们描述了一种方法，用于分离和培养平滑肌细胞来自成年大鼠（或小鼠）的上级肠系膜动脉。通过酶促解离获得动脉肌细胞并在原代培养物中建立。培养的细胞保留平滑肌特异性α-肌动蛋白的表达和对激动剂的生理反应。培养的动脉肌细胞（从野生型或转基因动物制备）提供了一个有用的模型，用于研究在细胞和亚细胞水平上调节广泛的血管平滑肌反应。质粒、RNA干扰和反义寡脱氧核苷酸可以容易地引入细胞中以改变蛋白质表达。荧光染料也可以被引入到可视化的各种活动，其中一些可能是特定的血管平滑肌细胞。该方案需要连续2天每天约3小时才能完成。
In this protocol, we describe a method for isolation and culture of smooth muscle cells derived from the adult rat (or mouse) superior mesenteric artery. Arterial myocytes are obtained by enzymatic dissociation and established in primary culture. The cultured cells retain expression of smooth muscle-specific alpha-actin and physiological responses to agonists. Cultured arterial myocytes (prepared from wild-type or transgenic animals) provide a useful model for studying the regulation of a wide range of vascular smooth muscle responses at the cellular and subcellular levels. Plasmids, RNA interference and antisense oligodeoxynucleotides can be readily introduced into the cells to alter protein expression. Fluorescent dyes can also be introduced to visualize a variety of activities, some of which may be specific to vascular smooth muscle cells. This protocol requires about 3 h on each of 2 consecutive days to complete.