Immortalized human choroid plexus endothelial cells enable an advanced endothelial-epithelial two-cell type in vitro model of the choroid plexus

Walter Muranyi; Christian Schwerk; Rosanna Herold; Carolin Stump-Guthier; Marko Lampe; Petra Fallier-Becker; Christel Weiß; Carsten Sticht; Hiroshi Ishikawa; Horst Schroten

doi:10.1016/j.isci.2022.104383

Immortalized human choroid plexus endothelial cells enable an advanced endothelial-epithelial two-cell type in vitro model of the choroid plexus

iScience. 2022 May 10;25(6):104383. doi: 10.1016/j.isci.2022.104383. eCollection 2022 Jun 17.

Authors

Walter Muranyi^{1

2}, Christian Schwerk^{1

2}, Rosanna Herold^{1

2}, Carolin Stump-Guthier^{1

2}, Marko Lampe³, Petra Fallier-Becker⁴, Christel Weiß⁵, Carsten Sticht⁶, Hiroshi Ishikawa⁷, Horst Schroten^{1

2}

Affiliations

¹ Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
² European Center for Angioscience, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
³ Advanced Light Microscopy Facility, European Molecular Biology Laboratory, Heidelberg, Germany.
⁴ Institute of Pathology and Neuropathology, University of Tübingen, Tübingen, Germany.
⁵ Department of Medical Statistics and Biomathematics, University Medical Center Mannheim, Heidelberg University, Heidelberg, Germany.
⁶ Core Facility Next Generation Sequencing, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
⁷ Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan.

Abstract

The choroid plexus (CP) is a highly vascularized structure containing endothelial and epithelial cells located in the ventricular system of the central nervous system (CNS). The role of the fenestrated CP endothelium is under-researched and requires the generation of an immortalized CP endothelial cell line with preserved features. Transduction of primary human CP endothelial cells (HCPEnC) with the human telomerase reverse transcriptase (hTERT) resulted in immortalized HCPEnC (iHCPEnC), which grew as monolayer with contact inhibition, formed capillary-like tubes in Matrigel, and showed no colony growth in soft agar. iHCPEnC expressed pan-endothelial markers and presented characteristic plasmalemma vesicle-associated protein-containing structures. Cultivation of iHCPEnC and human epithelial CP papilloma (HIBCPP) cells on opposite sides of cell culture filter inserts generated an in vitro model with a consistently enhanced barrier function specifically by iHCPEnC. Overall, iHCPEnC present a tool that will contribute to the understanding of CP organ functions, especially endothelial-epithelial interplay.

Keywords: Biological sciences; Biological sciences research methodologies; Biology experimental methods; Cell biology.