Establishment of a new conditionally immortalized human brain microvascular endothelial cell line retaining an in vivo blood–brain barrier function
Version of Record online: 10 MAY 2010
Copyright © 2010 Wiley-Liss, Inc.
Journal of Cellular Physiology
Volume 225, Issue 2, pages 519–528, November 2010
How to Cite
Sano, Y., Shimizu, F., Abe, M., Maeda, T., Kashiwamura, Y., Ohtsuki, S., Terasaki, T., Obinata, M., Kajiwara, K., Fujii, M., Suzuki, M. and Kanda, T. (2010), Establishment of a new conditionally immortalized human brain microvascular endothelial cell line retaining an in vivo blood–brain barrier function. J. Cell. Physiol., 225: 519–528. doi: 10.1002/jcp.22232
- Issue online: 23 AUG 2010
- Version of Record online: 10 MAY 2010
- Manuscript Accepted: 21 APR 2010
- Manuscript Received: 17 MAY 2009
- Japan Society for the Promotion of Science, Tokyo, Japan. Grant Numbers: 18390259, 19790610
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. J. Cell. Physiol. 225: 519–528, 2010. © 2010 Wiley-Liss, Inc.