| Isolation and Cultivation of Primary Brain Endothelial Cells from Adult Mice
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Author:
Date:
2017-05-20
[Abstract] Brain endothelial cells are the major building block of the blood-brain barrier. To study the role of brain endothelial cells in vitro, the isolation of primary cells is of critical value. Here, we describe a protocol in which vessel fragments are isolated from adult mice. After density centrifugation and mild digestion of the fragments, outgrowing endothelial cells are selected by puromycin treatment and grown to confluence within one week.
[摘要] 脑内皮细胞是血脑屏障的主要组成部分。 为了研究脑内皮细胞在体外的作用，原代细胞的分离是临床价值。 在这里，我们描述了一种方案，其中血管片段与成年小鼠分离。 在密度离心和片段的温和消化后，通过嘌呤霉素处理选择出生长的内皮细胞，并在一周内生长至汇合。
【背景】血脑屏障保护大脑免受细胞和物质的不受控制的侵入。 这主要是由脑内皮细胞形成的屏障组成的紧密和粘附连接限制细胞旁运输。
该方案被开发以克服维持其关键特征的小鼠脑内皮细胞系的有限可用性，例如表达足够量的紧密连接蛋白如闭合蛋白，ZO-1或紧密连接蛋白-5以诱导高跨内皮细胞抗性。
此外，从内含基因修饰的小鼠中分离脑内皮细胞可以在体外研究基因特异性功能。
使用这种方法，我们以前补充了体内研究，证明NF-κB信号在脑内皮细胞中维持正常血脑屏障功能的重要性（Ridder等，2015）。
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| Make Human Umbilical Vein Endothelial Cells from Cords
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Author:
Date:
2012-04-20
[Abstract] Human umbilical vein endothelial cells (HUVEC) can be isolated from normal human umbilical veins, and are responsive to cytokine stimulation by expressing cell adhesion molecules. These cell systems are commonly used for physiological and pharmacological investigations, such as macromolecule transport, blood coagulation, and angiogenesis. This protocol describes the generation of HUVECs.
[摘要] 人脐静脉内皮细胞(HUVEC)是从人脐静脉分离的，在表达细胞粘着分子会迅速应答细胞因子的刺激。这个细胞系统通常用于生理学和药理学的研究，例如高分子运输，血液凝固和血管发生。
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| Isolation of Endothelial Cells from Mice
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Author:
Date:
2012-04-20
[Abstract] Endothelial cells line the entire circulatory system, from the heart to the smallest capillary. In this protocol, attaining and maintaining vitality of the organ is critical. Minimizing the processing time between removing the organ and beginning the digestions will improve yields as cells are more viable and are not under hypoxic conditions for long. For this protocol to be most effective, mice should be 5-6 weeks of age. Older mice result in lower yields of endothelial cells. In our hands, as few as 5 mice can result in good yields.
[摘要] 内皮细胞分布于整个循环系统，从心脏到最小的毛细血管。在这个步骤中，气管的活力很非常重要。取出器官和开始消化之间的时间越短将提高细胞更有活力的产出及不在含氧量低的条件下太久。小鼠的数量需要：小鼠应该是5-6周大小。年龄大的小鼠内皮细胞的产出会少些。最少5个小鼠可以得到很好的产出。
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