Abstract
Understanding blood-brain barrier function under physiological and pathophysiological conditions is critical for the development of new therapeutic strategies that hold the promise to enhance brain drug delivery, improve brain protection, and treat brain disorders. However, studying the human blood-brain barrier function is challenging. Thus, there is a critical need for appropriate models. In this regard, brain capillaries isolated from human brain tissue represent a unique tool to study barrier function as close to the human in vivo situation as possible. Here, we describe an optimized protocol to isolate capillaries from human brain tissue at a high yield and with consistent quality and purity. Capillaries are isolated from fresh human brain tissue using mechanical homogenization, density-gradient centrifugation, and filtration. After the isolation, the human brain capillaries can be used for various applications including leakage assays, live cell imaging, and immune-based assays to study protein expression and function, enzyme activity, or intracellular signaling. Isolated human brain capillaries are a unique model to elucidate the regulation of the human blood-brain barrier function. This model can provide insights into central nervous system (CNS) pathogenesis, which will help the development of therapeutic strategies for treating CNS disorders.
Document Type
Article
Publication Date
9-12-2018
Digital Object Identifier (DOI)
https://doi.org/10.3791/57346
Funding Information
We thank and acknowledge Dr. Peter Nelson and Sonya Anderson at the UK-ADC Brain Tissue Bank for providing all human brain tissue samples (NIH grant number: P30 AG028383 from the National Institute on Aging).
This project was supported by grant number 1 R01 NS079507 from the National Institute of Neurological Disorders and Stroke (to B.B.) and by grant number 1 R01AG039621 from the National Institute on Aging (to A.M.S.H.).
Related Content
The video component of this article can be found at https://www.jove.com/video/57346/.
Repository Citation
Hartz, Anika M. S.; Schulz, Julia A.; Sokola, Brent S.; Edelmann, Stephanie E.; Shen, Andrew N.; Rempe, Ralf G.; Zhong, Yu; Seblani, Nader El; and Bauer, Bjoern, "Isolation of Cerebral Capillaries from Fresh Human Brain Tissue" (2018). Sanders-Brown Center on Aging Faculty Publications. 111.
https://uknowledge.uky.edu/sbcoa_facpub/111
Materials List
Notes/Citation Information
Published in Journal of Visualized Experiments, issue 139, e57346, p. 1-12.
Copyright © 2018 Journal of Visualized Experiments
The copyright holder has granted the permission for posting the article here.