Abstract
AbstractObjectivesThe breakdown of the blood–brain barrier (BBB) has been considered to be a key step in the disease process of a number of neuroimmunological disorders. Although severalin vitroBBBmodels derived from human tissues have been established, no human conditionally immortalizedin vitroBBBmodels using a temperature‐sensitiveSV40‐Tantigen (tsA58) and human telomerase reverse transcriptase (hTERT) have ever been reported. In the present study, we established a new human brain microvascular endothelial cell line harboringtsA58andhTERTgenes, and extensively characterized this new model.MethodsTY08 cells, derived from the humanBBBand harboringtsA58, were infected with retroviruses possessinghTERTgenes. We examined whether this new model retains its barrier‐specific nature, independent of the passage number.ResultsThe obtained endothelial cell line, termedTY09, proliferated well under the permissive temperature and stopped growing under the non‐permissive temperature, despite the acquisition ofhTERTas an additional immortalizing gene. Even with a high‐passage number, the cells maintained a spindle‐shaped morphology, the expression of the vonWillebrand factor, tight junction proteins and transporters. Furthermore, we carried out a transendothelial transport study forTY09 cells and hCMEC/D3 cells, thereby proving that both cell lines have almost the same nature with respect to transcellular permeability of various hydrophilic and hydrophobic substances.ConclusionsThe new stable conditionally immortalizedTY09 cells, retaining thein vivoBBBfunctions, should facilitate the performance of future studies for determining the pathophysiology of various neuroimmunological diseases.
Published Version
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