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A Triple Primary Cell Culture Model of the Human Blood-Brain Barrier for Studying Ischemic Stroke In Vitro.
Fattakhov, Nikolai; Torices, Silvia; Becker, Sarah; Teglas, Timea; Naranjo, Oandy; Toborek, Michal.
Afiliación
  • Fattakhov N; Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami; nfattakhov@miami.edu.
  • Torices S; Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami.
  • Becker S; Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami.
  • Teglas T; Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami.
  • Naranjo O; Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami.
  • Toborek M; Department of Biochemistry and Molecular Biology, Miller School of Medicine, University of Miami; Institute of Physiotherapy and Health Sciences, The Jerzy Kukuczka Academy of Physical Education; mtoborek@med.miami.edu.
J Vis Exp ; (188)2022 10 06.
Article en En | MEDLINE | ID: mdl-36282703
Ischemic stroke is a major cause of death and disability worldwide with limited therapeutic options. The neuropathology of ischemic stroke is characterized by an interruption in blood supply to the brain leading to cell death and cognitive dysfunction. During and after ischemic stroke, blood-brain barrier (BBB) dysfunction facilitates injury progression and contributes to poor patient recovery. Current BBB models primarily include endothelial monocultures and double co-cultures with either astrocytes or pericytes. Such models lack the ability to fully imitate a dynamic brain microenvironment, which is essential for cell-to-cell communication. Additionally, commonly used BBB models often contain immortalized human endothelial cells or animal-derived (rodent, porcine, or bovine) cell cultures that pose translational limitations. This paper describes a novel well-insert-based BBB model containing only primary human cells (brain microvascular endothelial cells, astrocytes, and brain vascular pericytes) enabling the investigation of ischemic brain injury in vitro. The effects of oxygen-glucose deprivation (OGD) on barrier integrity were assessed by passive permeability, transendothelial electrical resistance (TEER) measurements,and direct visualization of hypoxic cells. The presented protocol offers a distinct advantage inmimicking the intercellular environment of the BBB in vivo, serving as a more realistic in vitro BBB model for developing new therapeutic strategies in the setting of ischemic brain injury.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Lesiones Encefálicas / Accidente Cerebrovascular Isquémico Tipo de estudio: Guideline Límite: Animals / Humans Idioma: En Revista: J Vis Exp Año: 2022 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: Lesiones Encefálicas / Accidente Cerebrovascular Isquémico Tipo de estudio: Guideline Límite: Animals / Humans Idioma: En Revista: J Vis Exp Año: 2022 Tipo del documento: Article Pais de publicación: Estados Unidos