They were reported to have an apparently normal diploid human karyotype They do not show adhesion-independent growth in soft agar but form capillary structures in Matrigel, a characteristic property of cultured endothelium. The hCMEC/D3 cells form a contact-inhibited monolayer of elongated cells on collagen type I or type IV.
In the first passage, cells were sequentially immortalized by lentiviral vector transduction with the catalytic subunit of human telomerase (hTERT) and SV40 large T antigen, following which CEC were selectively isolated by limited dilution cloning, and clones were extensively characterized for brain endothelial phenotype The primary isolate was enriched in CECs. The hCMEC/D3 cell line was derived from human temporal lobe microvessels isolated from tissue excised during surgery for control of epilepsy. Here we review the suitability of this cell line as a human BBB model for pathogenic and drug transport studies and we critically consider its advantages and limitations.ĭerivation and selection of hCMEC/D3 cells Indeed, since the development of this cell line in 2005 over 100 studies on different aspects of cerebral endothelial biology and pharmacology have been published. The brain microvascular endothelial cell line hCMEC/D3 represents one such model of the human BBB that can be easily grown and is amenable to cellular and molecular studies on pathological and drug transport mechanisms with relevance to the central nervous system (CNS). While primary human CECs would ideally be the model of choice, the paucity of available fresh human cerebral tissue makes wide-scale studies impractical. Since the first attempts in the 1970s to isolate cerebral microvessel endothelial cells (CECs) in order to model the blood–brain barrier (BBB) in vitro, the need for a human BBB model that closely mimics the in vivo phenotype and is reproducible and easy to grow, has been widely recognized by cerebrovascular researchers in both academia and industry.
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