Development and characterization of a novel human in vitro blood-nerve barrier model using primary endoneurial endothelial cells.
There are phenotypic and functional differences between vascular endothelium from different tissues and between microvascular and macrovascular endothelial cells (ECs) from the same tissue. Relatively little is known about the human blood-nerve barrier (BNB). We report the development of an in vitro BNB model using primary human endoneurial ECs freshly isolated and purified from decedent sciatic nerves via endoneurial stripping, connective tissue enzymatic digestion, and density centrifugation. Primary human endoneurial ECs are spindle shaped and contact inhibited. They rapidly differentiate to form capillary-like networks and microvessels, bind Ulex Europaeus Agglutinin 1 lectin, express von Willebrand factor, and endocytose acetylated low-density lipoprotein. They also express specific transport and cellular adhesion molecules and tight junction proteins, consistent with cells that form a highly restrictive endothelial barrier similar to the blood-brain barrier. When cultured on collagen-coated transwell inserts, the primary human endoneurial ECs develop an in vitro BNB with high transendothelial electrical resistances (160 Omega x cm(2); maximal 12 days after seeding) and low solute permeability coefficient to fluoresceinated high-molecular weight (70 kDa) dextran (2.75 x 10(-3) cm/minute). This in vitro BNB model retains essential known or expected characteristics of the human BNB and has many potential applications for studies of solute, macromolecule, microbial, virus, and leukocyte interactions with this highly specialized endothelial barrier.
Yosef
Nejla
N
Neuromuscular Immunopathology Research Laboratory, Department of Neurology, Baylor College of Medicine, Houston, Texas 77030-3411, USA.
Xia
Robin H
RH
Ubogu
Eroboghene E
EE
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000199:Actins / Q000378:metabolism; D001692:Biological Transport / Q000502:physiology; D055592:Biophysical Phenomena / Q000502:physiology; D001812:Blood-Brain Barrier / Q000502:physiology* / Q000648:ultrastructure; D049428:Blood-Nerve Barrier / Q000166:cytology*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D003911:Dextrans / Q000378:metabolism; D017097:Electric Impedance; D004558:Electric Stimulation / Q000379:methods; D042783:Endothelial Cells / Q000502:physiology* / Q000648:ultrastructure; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D008077:Lipoproteins, LDL / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D008954:Models, Biological; D018919:Neovascularization, Physiologic / Q000502:physiology; D010525:Peripheral Nerves / Q000166:cytology*; D037121:Plant Lectins / Q000378:metabolism; D012584:Sciatic Nerve / Q000166:cytology; D019108:Tight Junctions / Q000378:metabolism / Q000648:ultrastructure; D005723:gamma-Glutamyltransferase / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism
C541116:ACTA2 protein, human; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000199:Actins; D015815:Cell Adhesion Molecules; D003911:Dextrans; D008077:Lipoproteins, LDL; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D037121:Plant Lectins; C036850:Ulex europaeus lectins; D014841:von Willebrand Factor; D005723:gamma-Glutamyltransferase