Procedure for the Isolation of Endothelial Cells from Human Cerebral Arteriovenous Malformation (cAVM) Tissues.
Procedure for the Isolation of Endothelial Cells from Human Cerebral Arteriovenous Malformation (cAVM) Tissues.
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从人脑动静脉畸形 (cAVM) 组织中分离内皮细胞的程序
DOI：
10.3389/fncel.2018.00030
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发表时间：
2018
影响因子：
5.3
通讯作者：
Zhao YL
中科院分区：
文献类型：
--
作者：
Hao Q;Chen XL;Ma L;Wang TT;Hu Y;Zhao YL
在这项研究中，我们成功地建立了一种稳定的从人脑动静脉畸形（cAVM）组织中分离内皮细胞（ECs）的方法。尽管人类cAVM组织中ECs的数量较少，但它们在cAVM的表现和发展以及出血性卒中和血栓形成中发挥着重要作用。为了表征和了解人cAVM中内皮细胞的生物学特性，分离和纯化这些细胞的方法是必要的。我们已经开发了这种方法，以可靠地从cavm中获得纯ECs群体。为了获得纯细胞群，对cAVM组织进行机械和酶消化，然后用特定细胞抗原的抗体标记单个cAVM- ecs悬液，并通过流式细胞术选择。纯化后的内皮细胞采用免疫染色特异性抗原检测，并用于研究不同的细胞机制。与从组织中分离内皮细胞的不同方法相比，我们可以自信地从cavm中分离出内皮细胞，并且收获的cavm -内皮细胞的数量几乎与血管成分中的数量相似。除了优化从人cAVM组织中分离内皮细胞的方案外，该方案还可用于从其他人类神经血管病变组织中分离内皮细胞。根据组织的不同，整个过程可以在大约20天内完成。
In this study, we successfully established a stable method for the isolation of endothelial cells (ECs) from human cerebral arteriovenous malformation (cAVM) tissues. Despite human cAVM tissues having a minor population of ECs, they play an important role in the manifestation and development of cAVM as well as in hemorrhagic stroke and thrombogenesis. To characterize and understand the biology of ECs in human cAVM (cAVM-ECs), methods for the isolation and purification of these cells are necessary. We have developed this method to reliably obtain pure populations of ECs from cAVMs. To obtain pure cell populations, cAVM tissues were mechanically and enzymatically digested and the resulting single cAVM-ECs suspensions were then labeled with antibodies of specific cell antigens and selected by flow cytometry. Purified ECs were detected using specific makers of ECs by immunostaining and used to study different cellular mechanisms. Compared to the different methods of isolating ECs from tissues, we could isolate ECs from cAVMs confidently, and the numbers of cAVM-ECs harvested were almost similar to the amounts present in vessel components. In addition to optimizing the protocol for isolation of ECs from human cAVM tissues, the protocol could also be applied to isolate ECs from other human neurovascular-diseased tissues. Depending on the tissues, the whole procedure could be completed in about 20 days.
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影响因子：
14.8
作者：
Levenberg, Shulamit;Ferreira, Lino S.;Chen-Konak, Limor;Kraehenbuehl, Thomas P.;Langer, Robert
通讯作者：
Langer, Robert
影响因子：
8.3
作者：
Ferreira, Raquel;Santos, Tiago;Hofman, Florence M.
通讯作者：
Hofman, Florence M.
影响因子：
3.7
作者：
van Beijnum, Judy R.;van der Linden, Edith;Griffioen, Arjan W.
通讯作者：
Griffioen, Arjan W.
影响因子：
2.5
作者：
Zhang, Hai-Feng;Liang, Guo-Biao;Xu, Zai-Hua
通讯作者：
Xu, Zai-Hua
影响因子：
2
作者：
Young, Adam M. H.;Teo, Mario;St George, E. Jerome
通讯作者：
St George, E. Jerome