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

doi:10.1016/j.xpro.2022.101816

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

STAR Protoc. 2022 Nov 4;3(4):101816. doi: 10.1016/j.xpro.2022.101816. eCollection 2022 Dec 16.

Authors

Rosanna Herold¹, Julia Borkowski¹, Christian Schwerk¹, Carolin Stump-Guthier¹, Marko Lampe², Hiroshi Ishikawa³, Walter Muranyi⁴, Horst Schroten¹

Affiliations

¹ 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.
² European Molecular Biology Laboratory, Advanced Light Microscopy Core Facility, 69117 Heidelberg, Germany.
³ Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
⁴ 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. Electronic address: walter.muranyi@medma.uni-heidelberg.de.

Abstract

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).

Keywords: Cell biology; Cell culture; Molecular biology; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blood-Brain Barrier
Cell Count
Choroid Plexus*
Endothelial Cells*
Epithelial Cells
Humans