Organotypic brain slice cultures as a model to study angiogenesis of brain vessels
Organotypic brain slice cultures as a model to study angiogenesis of brain vessels
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DOI：
10.3389/fcell.2015.00052
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发表时间：
2015-09-02
影响因子：
5.5
通讯作者：
Humpel, Christian
中科院分区：
文献类型：
--
作者：
Hutter-Schmid, Bianca;Kniewallner, Kathrin M.;Humpel, Christian
脑血管是大脑中向神经元传递能量和底物的最重要结构。脑血管由内皮细胞、周细胞和星形胶质细胞之间复杂的相互作用组成，控制底物进入大脑。脑血管损伤和血管损伤是在不同的神经退行性疾病（包括例如阿尔茨海默病）中观察到的一般病理学。为了研究脑血管的重塑，需要开发简单的 3 维体外系统。小鼠器官型脑切片为探索复杂三维结构中脑血管的血管生成效应提供了有效的工具。在这里，我们展示了可以从出生后和成年小鼠大脑的 110 μm 厚切片中培养器官型脑切片。对血管进行层粘连蛋白和 IV 型胶原蛋白的免疫组织化学染色。共染色是可视化脑内皮细胞与这些血管中的周细胞和星形胶质细胞相互作用的合适方法。不同的外源刺激，例如成纤维细胞生长因子-2或血管内皮生长因子，分别诱导血管生成或再生长。体温过高或酸中毒会降低器官切片中的血管密度。总之，器官型脑切片表现出强大的血管网络，可用于研究 3 维体外系统中脑血管的重塑和血管生成。
Brain vessels are the most important structures in the brain to deliver energy and substrates to neurons. Brain vessels are composed of a complex interaction between endothelial cells, pericytes, and astrocytes, controlling the entry of substrates into the brain. Damage of brain vessels and vascular impairment are general pathologies observed in different neurodegenerative disorders including e.g., Alzheimer's disease. In order to study remodeling of brain vessels, simple 3-dimensional in vitro systems need to be developed. Organotypic brain slices of mice provide a potent tool to explore angiogenic effects of brain vessels in a complex 3-dimensional structure. Here we show that organotypic brain slices can be cultured from 110 mu m thick sections of postnatal and adult mice brains. The vessels are immunohistochemically stained for laminin and collagen IV. Co-stainings are an appropriate method to visualize interaction of brain endothelial cells with pericytes and astrocytes in these vessels. Different exogenous stimuli such as fibroblast growth factor-2 or vascular endothelial growth factor induce angiogenesis or re-growth, respectively. Hyperthermia or acidosis reduces the vessel density in organotypic slices. In conclusion, organotypic brain slices exhibit a strong vascular network which can be used to study remodeling and angiogenesis of brain vessels in a 3-dimensional in vitro system.