Protocol for the setup and use of a human choroid plexus endothelial-epithelial two-cell-type in vitro model

Rosanna Herold; Julia Borkowski; Christian Schwerk; Carolin Stump-Guthier; Marko Lampe; Hiroshi Ishikawa; Walter Muranyi; Horst Schroten

STAR Protocols (Dec 2022)

Protocol for the setup and use of a human choroid plexus endothelial-epithelial two-cell-type in vitro model

Rosanna Herold,
Julia Borkowski,
Christian Schwerk,
Carolin Stump-Guthier,
Marko Lampe,
Hiroshi Ishikawa,
Walter Muranyi,
Horst Schroten

Affiliations

Rosanna Herold: Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany; European Center for Angioscience, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany
Julia Borkowski: Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany; European Center for Angioscience, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany
Christian Schwerk: Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany; European Center for Angioscience, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany
Carolin Stump-Guthier: Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany; European Center for Angioscience, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany
Marko Lampe: European Molecular Biology Laboratory, Advanced Light Microscopy Core Facility, 69117 Heidelberg, Germany
Hiroshi Ishikawa: Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
Walter Muranyi: Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany; European Center for Angioscience, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany; Corresponding author
Horst Schroten: Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany; European Center for Angioscience, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany

Journal volume & issue: Vol. 3, no. 4
p. 101816

Abstract

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Summary: Choroid plexus, located in brain ventricles, is the site of blood-cerebrospinal fluid barrier that contains endothelial cells and an epithelial monolayer separated by stroma. We established a two-cell-type model of the human choroid plexus consisting of immortalized endothelial cells (iHCPEnC) and epithelial papilloma (HIBCPP) cells grown on opposite sides of filter supports. In this protocol, we describe the preparation of this model, the measurement of transepithelial electrical resistance (TEER), and immunofluorescence imaging-based analysis to determine the barrier function.For complete details on the use and execution of this protocol, please refer to Muranyi et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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