title: Establishment of a new conditionally immortalized human brain microvascular endothelial cell line retaining an in vivo blood-brain barrier function.
abstract: The breakdown of the blood-brain barrier (BBB) has been considered to be a key step in the disease process of a number of neurological disorders such as cerebral ischemia and Alzheimer's disease. Many in vitro BBB models derived from animal tissues have been established to elucidate the mechanism of BBB insufficiency. However, only a few human immortalized in vitro BBB models have been reported. In the present study, a temperature-sensitive SV40-T antigen was introduced to immortalize cells using a retrovirus to obtain a better human in vitro BBB model which sustains physiological properties. This endothelial cell (EC) line, termed TY08, showed a spindle-shaped morphology. The cells expressed all key tight junctional proteins, such as occludin, claudin-5, zonula occludens (ZO)-1 and ZO-2 at their cell-to-cell boundaries, and had low permeability to inulin across its monolayer. The cells also expressed various influx and efflux transporters and exhibited the functional expression of p-glycoprotein. Furthermore, the TY08 cells grew and proliferated well under the permissive temperature and stopped growing under the non-permissive temperature to serve as physiological ECs forming the BBB. Thus, conditionally immortalized TY08 cells retaining the in vivo BBB functions should facilitate analyses for determining the pathophysiology of various neurological diseases.
authors.affiliation: Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi, Japan.
authors.forename: Yasuteru
authors.initials: Y
authors.lastname: Sano
authors.forename: Fumitaka
authors.initials: F
authors.lastname: Shimizu
authors.forename: Masaaki
authors.initials: M
authors.lastname: Abe
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Maeda
authors.forename: Yoko
authors.initials: Y
authors.lastname: Kashiwamura
authors.forename: Sumio
authors.initials: S
authors.lastname: Ohtsuki
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Terasaki
authors.forename: Masuo
authors.initials: M
authors.lastname: Obinata
authors.forename: Koji
authors.initials: K
authors.lastname: Kajiwara
authors.forename: Masami
authors.initials: M
authors.lastname: Fujii
authors.forename: Michiyasu
authors.initials: M
authors.lastname: Suzuki
authors.forename: Takashi
authors.initials: T
authors.lastname: Kanda
mesh_terms: D001812:Blood-Brain Barrier / Q000502:physiology*; D001921:Brain / Q000098:blood supply*; D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology*; D020869:Gene Expression Profiling; D006309:Hearing; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019108:Tight Junctions
chemical_list: D008565:Membrane Proteins