Abstract
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.
Highlights
Cerebral arteriovenous malformations are vascular lesions characterized by abnormal arteries and venous entanglement, which divert blood directly from the arteries to the venous circulation, instead of capillaries. cerebral arteriovenous malformation (cAVM) are rare and occur sporadically, which may be associated with genetic disorders (Consoli et al, 2013; Novakovic et al, 2013; Gross and Du, 2014; Young et al, 2015; Figure 1)
Using the protocol we developed, isolated cAVM-endothelial cells (ECs) grew into separate colonies, with morphologies of round borders and cobblestone appearance
All primary ECs established from cAVMs were viable for a long period and expressed specific cell receptors, such as CD31, CD34, and von Willebrand factor (vWF)
Summary
Cerebral arteriovenous malformations (cAVMs) are vascular lesions characterized by abnormal arteries and venous entanglement, which divert blood directly from the arteries to the venous circulation, instead of capillaries. cAVMs are rare and occur sporadically, which may be associated with genetic disorders (Consoli et al, 2013; Novakovic et al, 2013; Gross and Du, 2014; Young et al, 2015; Figure 1). A method to isolate and purify ECs from cAVM tissues is essential. These single-cell suspensions were labeled with antibodies and isolated and purified through FACS (Figures 2, 3).
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
