title: Co-culture model consisting of human brain microvascular endothelial and peripheral blood mononuclear cells.
abstract: 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.
authors.affiliation: Department of Microbiology and Immunology, Center for Molecular Virology and Translational Neuroscience, Institute for Molecular Medicine and Infectious Disease, Drexel University College of Medicine, 245 N. 15th Street, MS #1013A, Philadelphia, PA 19102, USA. Electronic address: ms996@drexel.edu.
authors.forename: Marianne
authors.initials: M
authors.lastname: Strazza
authors.affiliation: Department of Microbiology and Immunology, Center for Molecular Virology and Translational Neuroscience, Institute for Molecular Medicine and Infectious Disease, Drexel University College of Medicine, 245 N. 15th Street, MS #1013A, Philadelphia, PA 19102, USA. Electronic address: mem446@drexel.edu.
authors.forename: Monique E
authors.initials: ME
authors.lastname: Maubert
authors.affiliation: Department of Microbiology and Immunology, Center for Molecular Virology and Translational Neuroscience, Institute for Molecular Medicine and Infectious Disease, Drexel University College of Medicine, 245 N. 15th Street, MS #1013A, Philadelphia, PA 19102, USA. Electronic address: vpirrone@drexelmed.edu.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Pirrone
authors.affiliation: Department of Microbiology and Immunology, Center for Molecular Virology and Translational Neuroscience, Institute for Molecular Medicine and Infectious Disease, Drexel University College of Medicine, 245 N. 15th Street, MS #1013A, Philadelphia, PA 19102, USA; Sidney Kimmel Cancer Center, Thomas Jefferson University, Bluemle Life Sciences Building, 233 South 10th Street Suite 1050, Philadelphia, PA 19107, USA. Electronic address: bwigdahl@drexelmed.edu.
authors.forename: Brian
authors.initials: B
authors.lastname: Wigdahl
authors.affiliation: Department of Microbiology and Immunology, Center for Molecular Virology and Translational Neuroscience, Institute for Molecular Medicine and Infectious Disease, Drexel University College of Medicine, 245 N. 15th Street, MS #1013A, Philadelphia, PA 19102, USA. Electronic address: mnonnema@drexelmed.edu.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Nonnemacher
mesh_terms: D001812:Blood-Brain Barrier / Q000502:physiology; D002448:Cell Adhesion; D002460:Cell Line; D002465:Cell Movement; D002470:Cell Survival; D018920:Coculture Techniques; D003094:Collagen; D003911:Dextrans; D042783:Endothelial Cells / Q000502:physiology; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D006801:Humans; D006854:Hydrocortisone / Q000378:metabolism / Q000633:toxicity; D007963:Leukocytes, Mononuclear / Q000502:physiology; D055806:Microvessels / Q000502:physiology; D011138:Polytetrafluoroethylene; D054457:Tissue Scaffolds
chemical_list: D003911:Dextrans; C015219:fluorescein isothiocyanate dextran; D011138:Polytetrafluoroethylene; D003094:Collagen; D016650:Fluorescein-5-isothiocyanate; D006854:Hydrocortisone
keywords: Blood–brain barrier; Brain microvascular endothelial cells; Peripheral blood mononuclear cells; hCMEC/D3 cell line