Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification.
Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification.
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DOI：
10.3791/56126
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发表时间：
2017-11-20
期刊：
影响因子：
--
通讯作者：
Macrae VE
中科院分区：
文献类型：
--
作者：
Lin C;Zhu D;Markby G;Corcoran BM;Farquharson C;Macrae VE
钙化性主动脉瓣疾病（CAVD）的特征是主动脉瓣叶进行性增厚。这是一种常见于老年人和终末期肾病（ESRD）患者的疾病，他们通常患有高磷酸盐血症和高钙血症。目前，没有药物治疗可以阻止其进展。这种病理过程的机制尚不清楚。主动脉瓣叶由主动脉瓣尖外表面上的一薄层瓣膜内皮细胞（VEC）组成，瓣膜间质细胞（VIC）夹在VEC之间。使用大鼠模型能够基于患有高磷酸盐血症和高钙血症的患者的体内生理病理学血清磷酸盐（Pi）和钙（Ca）水平进行异位钙化的体外研究。所述方案详细描述了纯大鼠维克群的分离，如维克标志物：α-平滑肌肌动蛋白（α-SMA）波形蛋白和组织生长因子β（TGFβ）1和2的表达所示，以及不存在分化簇（CD）31（一种VEC标志物）。通过扩大这些VIC，可以进行生化，遗传和成像研究，以研究和揭示CAVD基础的关键介质。
Calcific aortic valve disease (CAVD) is characterized by the progressive thickening of the aortic valve leaflets. It is a condition frequently found in the elderly and end-stage renal disease (ESRD) patients, who commonly suffer from hyperphosphatemia and hypercalcemia. At present, there are no medication therapies that can stop its progression. The mechanisms that underlie this pathological process remain unclear. The aortic valve leaflet is composed of a thin layer of valve endothelial cells (VECs) on the outer surfaces of the aortic cusps, with valve interstitial cells (VICs) sandwiched between the VECs. The use of a rat model enables the in vitro study of ectopic calcification based on the in vivo physiopathological serum phosphate (Pi) and calcium (Ca) levels of patients who suffer from hyperphosphatemia and hypercalcemia. The described protocol details the isolation of a pure rat VIC population as shown by the expression of VIC markers: alpha-smooth muscle actin (α-SMA) vimentin and tissue growth factor beta (TGFβ) 1 and 2, and the absence of cluster of differentiation (CD) 31, a VEC marker. By expanding these VICs, biochemical, genetic, and imaging studies can be performed to study and unravel the key mediators underpinning CAVD.
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