Procedure for the Isolation of Endothelial Cells from Human Cerebral Arteriovenous Malformation (cAVM) Tissues

Frontiers in Cellular Neuroscience

Yue Hu¹  Tong-Tong Wang¹  Yuan-Li Zhao²  Xiao-Lin Chen⁴  Li Ma⁴  Qiang Hao⁵ 

[1] Basic Medical Science Department, Capital Medical University, Beijing, China;Beijing Key Laboratory of Translational Medicine for Cerebrovascular Disease, Beijing, China;China National Clinical Research Center for Neurological Diseases, Beijing, China;Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China;Department of Neurosurgery, Peking University International Hospital, Peking University, Beijing, China;Stroke Center, Beijing Institute for Brain Disorders, Beijing, China;

关键词: endothelial cells;    cerebral arteriovenous malformation;    isolation approach;    purification;    angiogenesis;    in vitro culture;   

DOI  :  10.3389/fncel.2018.00030

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown