Establishment of a new conditionally immortalized human brain microvascular endothelial cell line retaining an in vivo blood-brain barrier function

Yasuteru Sano, Fumitaka Shimizu, Masaaki Abe, Toshihiko Maeda, Yoko Kashiwamura, Sumio Ohtsuki, Tetsuya Terasaki, Masuo Obinata, Koji Kajiwara, Masami Fujii, Michiyasu Suzuki, Takashi Kanda

Research output: Contribution to journalArticlepeer-review

86 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)519-528
Number of pages10
JournalJournal of Cellular Physiology
Volume225
Issue number2
DOIs
Publication statusPublished - 2010 Nov

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

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