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从大鼠大脑中动脉分离的平滑肌细胞的培养特性。

Characterization of culture from smooth muscle cells isolated from rat middle cerebral arteries.

机构信息

CICS-UBI - Centro de Investigação em Ciências da Saúde, University of Beira Interior, 6200-506 Covilhã, Portugal.

CICS-UBI - Centro de Investigação em Ciências da Saúde, University of Beira Interior, 6200-506 Covilhã, Portugal.

出版信息

Tissue Cell. 2020 Oct;66:101400. doi: 10.1016/j.tice.2020.101400. Epub 2020 Jun 27.

Abstract

Although human brain represents only 2% of the body mass, it uses around 20 % of the organism energy. Due to the brain's limited energy storage, the oxygen and glucose necessary to support brain functions depends on the correct blood supply. The main components of the arteries are smooth muscle cells, which are considered the main regulators of vascular tone and blood flow distribution. The information currently available on the functioning of the cerebral arteries and their cell constituents is extremely scarce. Thus, the aim of this work was to develop an in vitro model of smooth muscle cells derived from rat middle cerebral artery. Explants were collected from rat middle cerebral artery and adhered to collagen-coated culture dishes. Immunocytochemical analysis showed that the cells present in the culture expressed α-actin, a protein characteristic of the contractile phenotype of these cells. In addition, these cells did not express the endothelial marker, vWF. To evaluate the functionality of these cells the response to contractile agents, serotonin and noradrenaline, and to relaxing agent, sodium nitroprusside was determine by Planar Cell Surface Area analysis. Together the data obtained show that the cell culture obtained through the procedure described resulted in cells presenting the markers characteristic of smooth muscle cells and maintaining the usual contractile response, indicating that the cells obtained through this may be used as a model for characterization and study of functional behavior of the middle cerebral artery, as well as interaction studies between vascular and neuronal system.

摘要

尽管人脑仅占体重的 2%,但它却消耗了机体约 20%的能量。由于大脑的能量储存有限,因此支持大脑功能所需的氧气和葡萄糖依赖于正确的血液供应。动脉的主要成分是平滑肌细胞,平滑肌细胞被认为是血管张力和血流分布的主要调节者。目前关于脑动脉及其细胞成分功能的信息极为有限。因此,本研究旨在建立一种源自大鼠大脑中动脉的平滑肌细胞体外模型。从大鼠大脑中动脉采集培养物并贴附于涂有胶原蛋白的培养皿上。免疫细胞化学分析表明,培养物中的细胞表达α-肌动蛋白,这是这些细胞收缩表型的特征蛋白。此外,这些细胞不表达内皮标志物 vWF。为了评估这些细胞的功能,通过平面细胞表面积分析来测定其对收缩剂 5-羟色胺和去甲肾上腺素以及舒张剂硝普钠的反应。综合所得数据表明,通过所描述的程序获得的细胞培养物中存在具有平滑肌细胞特征性标志物的细胞,并保持了通常的收缩反应,这表明通过这种方法获得的细胞可用于对大脑中动脉的功能行为进行特征描述和研究,以及用于研究血管和神经元系统之间的相互作用。

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