Hi,
Would anyone know which generation of lentiviral vectors were used to generate hCMEC/d3 cell line?
Thank you,
Ecem
Dear Dr. Ecem Kaya
The hCMEC/D3 cell line was established by co-expressing hTERT with the large T antigen SV40 through a lentiviral vector system to immortalize primary human brain capillary endothelial cells (BCECs).
The hCMEC/D3 cell line was derived from human temporal lobe microvessels isolated from tissue excised during surgery for control of epilepsy. The primary isolate was enriched in cerebral microvessel endothelial cells (CECs). 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.
You may be interested in the paper attached below.
Article The hCMEC/D3 cell line as a model of the human blood brain barrier
For more detailed information, you may want to refer to the article attached below in the Materials and Methods section.
Sub-topic: Immortalization and selection of human brain endothelial cells.
Article Weksler BB, Subileau EA, Couraud POBlood-brain barrier-speci...
Regards,
Malcolm Nobre
Dear Malcolm Nobre
Thank you very much for your answer. I am aware of the paper you mentioned, however maybe it is me lacking the background on lentiviral vectors, I cannot understand or find the information on which generation of the lentiviral vectors that were used to generate this cell line (2nd? 3rd?).
Weksler et al paper uses DNA flap lentiviral vector and cites Zennou et al. 2000, Zennou et al 2001 in their methods section. I am hoping to find out a vector map so that I can make sure the vector(s) they use is third generation.
Thank you,
Ecem
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