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.
| Original language | English |
|---|---|
| Article number | e57346 |
| Journal | Journal of Visualized Experiments |
| Volume | 2018 |
| Issue number | 139 |
| DOIs | |
| State | Published - Sep 12 2018 |
Bibliographical note
Publisher Copyright:© 2018, Journal of Visualized Experiments. All rights reserved.
Funding
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). We thank Matt Hazzard and Tom Dolan, Information Technology Services, Academic Technology and Faculty Engagement, University of Kentucky for graphical assistance. This project was supported by grant number 1R01NS079507 from the National Institute of Neurological Disorders and Stroke (to B.B.) and by grant number 1R01AG039621 from the National Institute on Aging (to A.M.S.H.). The content is solely the responsibility of the authors and does not. 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). We thank Matt Hazzard and Tom Dolan, Information Technology Services, Academic Technology and Faculty Engagement, University of Kentucky for graphical assistance. This project was supported by grant number 1R01NS079507 from the National Institute of Neurological Disorders and Stroke (to B.B.) and by grant number 1R01AG039621 from the National Institute on Aging (to A.M.S.H.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Neurological Disorders and Stroke or the National Institute on Aging. The authors declare no competing financial interests.
| Funders | Funder number |
|---|---|
| UK-ADC | |
| National Institutes of Health (NIH) | P30 AG028383 |
| National Institutes of Health (NIH) | |
| National Institute on Aging | R01AG039621, 1R01NS079507 |
| National Institute on Aging | |
| Institute of Neurological Disorders and Stroke National Advisory Neurological Disorders and Stroke Council | |
| University of Kentucky |
Keywords
- Blood-brain barrier
- Brain capillaries
- Endothelial cells
- Human brain tissue
- Issue 139
- Neuroscience
- Neuroscience
- Neurovasculature
ASJC Scopus subject areas
- General Neuroscience
- General Chemical Engineering
- General Biochemistry, Genetics and Molecular Biology
- General Immunology and Microbiology