适用于分析细胞内钙和一氧化氮生物合成途径的大鼠冠状动脉内皮细胞分离培养高产方法。
High-Yield Method for Isolation and Culture of Endothelial Cells from Rat Coronary Blood Vessels Suitable for Analysis of Intracellular Calcium and Nitric Oxide Biosynthetic Pathways.
作者信息
Nistri Silvia, Mazzetti Luca, Failli Paola, Bani Daniele
机构信息
Department of Anatomy, Histology and Forensic Medicine, Section of Histology, University of Florence, Italy. V. le G. Pieraccini, 6, I-50139 Florence. Italy.; Department of Preclinical and Clinical Pharmacology, University of Florence, Italy. V. le G. Pieraccini, 6, I-50139 Florence. Italy.
出版信息
Biol Proced Online. 2002 Oct 28;4:32-37. doi: 10.1251/bpo31.
We describe here a method for isolating endothelial cells from rat heart blood vessels by means of coronary microperfusion with collagenase. This methods makes it possible to obtain high amounts of endothelial cells in culture which retain the functional properties of their in vivo counterparts, including the ability to uptake fluorescently-labeled acetylated low-density lipoproteins and to respond to vasoactive agents by modulating intracellular calcium and by upregulating intrinsic nitric oxide generation. The main advantages of our technique are: (i) good reproducibility, (ii) accurate sterility that can be maintained throughout the isolation procedure and (iii) high yield of pure endothelial cells, mainly due to microperfusion and temperature-controlled incubation with collagenase which allow an optimal distribution of this enzyme within the coronary vascular bed.
我们在此描述一种通过用胶原酶进行冠状动脉微灌注从大鼠心脏血管中分离内皮细胞的方法。该方法能够在培养中获得大量内皮细胞，这些细胞保留了其体内对应细胞的功能特性，包括摄取荧光标记的乙酰化低密度脂蛋白的能力，以及通过调节细胞内钙和上调内源性一氧化氮生成来对血管活性药物作出反应的能力。我们技术的主要优点是：（i）良好的可重复性，（ii）在整个分离过程中可保持的精确无菌性，以及（iii）高产量的纯内皮细胞，这主要归功于微灌注和与胶原酶的温度控制孵育，这使得该酶在冠状动脉血管床内实现最佳分布。
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