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Journal article
Co-culture model consisting of human brain microvascular endothelial and peripheral blood mononuclear cells
Journal of neuroscience methods, v 269, pp 39-45
30 Aug 2016
PMID: 27216631
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
•The immortalized human BMEC cell line, hCMEC/D3, in vitro blood-brain barrier model is useful in long-term functional studies.•hCMEC/D3 media components EBM-2 and hydrocortisone are cytotoxic to peripheral blood mononuclear cells (PBMCs).•Co-culture media excluding EBM-2 and hydrocortisone support hCMEC/D3 barrier function as well as continued PBMC viability.•Co-culture media can be used in assays of PBMC transmigration.
Numerous systems exist to model the blood–brain barrier (BBB) with the goal of understanding the regulation of passage into the central nervous system (CNS) and the potential impact of selected insults on BBB function. These models typically focus on the intrinsic cellular properties of the BBB, yet studies of peripheral cell migration are often excluded due to technical restraints.
This method allows for the study of in vitro cellular transmigration following exposure to any treatment of interest through optimization of co-culture conditions for the human brain microvascular endothelial cells (BMEC) cell line, hCMEC/D3, and primary human peripheral blood mononuclear cells (PBMCs).
hCMEC/D3 cells form functionally confluent monolayers on collagen coated polytetrafluoroethylene (PTFE) transwell inserts, as assessed by microscopy and tracer molecule (FITC-dextran (FITC-D)) exclusion. Two components of complete hCMEC/D3 media, EBM-2 base-media and hydrocortisone (HC), were determined to be cytotoxic to PBMCs. By combining the remaining components of complete hCMEC/D3 media with complete PBMC media a resulting co-culture media was established for use in hCMEC/D3—PBMC co-culture functional assays.
Through this method, issues of extensive differences in culture media conditions are resolved allowing for treatments and functional assays to be conducted on the two cell populations co-cultured simultaneously.
Described here is an in vitro co-culture model of the BBB, consisting of the hCMEC/D3 cell line and primary human PBMCs. The co-culture media will now allow for the study of exposure to potential insults to BBB function over prolonged time courses.
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Details
- Title
- Co-culture model consisting of human brain microvascular endothelial and peripheral blood mononuclear cells
- Creators
- Marianne Strazza - Drexel UniversityMonique E. Maubert - Drexel UniversityVanessa Pirrone - Drexel UniversityBrian Wigdahl - Drexel UniversityMichael R. Nonnemacher - Drexel University
- Publication Details
- Journal of neuroscience methods, v 269, pp 39-45
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:000380076300005
- Scopus ID
- 2-s2.0-84969814766
- Other Identifier
- 991019167712304721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemical Research Methods
- Neurosciences