Your browser doesn't support javascript.
loading
Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells.
Malik, Johid R; Modebelu, Ukamaka O; Fletcher, Courtney V; Podany, Anthony T; Scarsi, Kimberly K; Byrareddy, Siddappa N; Anand, Robbyn K; Buch, Shilpa; Sil, Susmia; Le, Jennifer; Bradley, John S; Brown, Ashley N; Sutar, Debapriya; Avedissian, Sean N.
Afiliación
  • Malik JR; Antiviral Pharmacology Laboratory, College of Pharmacy, University of Nebraska Medical Center, Omaha, Nebraska.
  • Modebelu UO; Antiviral Pharmacology Laboratory, College of Pharmacy, University of Nebraska Medical Center, Omaha, Nebraska.
  • Fletcher CV; Antiviral Pharmacology Laboratory, College of Pharmacy, University of Nebraska Medical Center, Omaha, Nebraska.
  • Podany AT; Division of Infectious Diseases, Department of Medicine, University of Nebraska Medical Center, Omaha, Nebraska.
  • Scarsi KK; Antiviral Pharmacology Laboratory, College of Pharmacy, University of Nebraska Medical Center, Omaha, Nebraska.
  • Byrareddy SN; Antiviral Pharmacology Laboratory, College of Pharmacy, University of Nebraska Medical Center, Omaha, Nebraska.
  • Anand RK; Division of Infectious Diseases, Department of Medicine, University of Nebraska Medical Center, Omaha, Nebraska.
  • Buch S; Department of Pharmacology & Experimental Neurosciences, University of Nebraska Medical Center, Omaha, Nebraska.
  • Sil S; Department of Chemistry, Iowa State University, Ames, Iowa.
  • Le J; Department of Pharmacology & Experimental Neurosciences, University of Nebraska Medical Center, Omaha, Nebraska.
  • Bradley JS; Department of Pharmacology & Experimental Neurosciences, University of Nebraska Medical Center, Omaha, Nebraska.
  • Brown AN; University of California San Diego, Skaggs School of Pharmacy and Pharmaceutical Science, La Jolla, California.
  • Sutar D; University of California San Diego School of Medicine, Department of Pediatrics, San Diego, California.
  • Avedissian SN; Institute for Therapeutic Innovation, College of Medicine, University of Florida, Orlando, Florida.
Curr Protoc ; 4(6): e1067, 2024 Jun.
Article en En | MEDLINE | ID: mdl-38857108
ABSTRACT
The blood-brain barrier (BBB) constitutes a crucial protective anatomical layer with a microenvironment that tightly controls material transit. Constructing an in vitro BBB model to replicate in vivo features requires the sequential layering of constituent cell types. Maintaining heightened integrity in the observed tight junctions during both the establishment and post-experiment phases is crucial to the success of these models. We have developed an in vitro BBB model that replicates the cellular composition and spatial orientation of in vivo BBB observed in humans. The experiment includes comprehensive procedures and steps aimed at enhancing the integration of the four-cell model. Departing from conventional in vitro BBB models, our methodology eliminates the necessity for pre-coated plates to facilitate cell adhesion, thereby improving cell visualization throughout the procedure. An in-house coating strategy and a simple yet effective approach significantly reduce costs and provides superior imaging of cells and corresponding tight junction protein expression. Also, our BBB model includes all four primary cell types that are structural parts of the human BBB. With its innovative and user-friendly features, our in-house optimized in vitro four-cell-based BBB model showcases novel methodology and provides a promising experimental platform for drug screening processes. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1 Coating and culture system Basic Protocol 2 Cell seeding and Transwell insert handling Basic Protocol 3 Assessment of model functionality.
Asunto(s)
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Barrera Hematoencefálica Límite: Humans Idioma: En Revista: Curr Protoc Año: 2024 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Barrera Hematoencefálica Límite: Humans Idioma: En Revista: Curr Protoc Año: 2024 Tipo del documento: Article Pais de publicación: Estados Unidos