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A Human Immortalized Cell-Based Blood–Brain Barrier Triculture Model: Development and Characterization as a Promising Tool for Drug−Brain Permeability Studies
dataset
posted on 2019-10-15, 13:47 authored by Ryo Ito, Kenta Umehara, Shota Suzuki, Keita Kitamura, Ken-ichi Nunoya, Yoshiyuki Yamaura, Haruo Imawaka, Saki Izumi, Naomi Wakayama, Takafumi Komori, Naohiko Anzai, Hidetaka Akita, Tomomi FurihataBrain
microvascular endothelial cells (BMEC), together with astrocytes
and pericytes, form the blood–brain barrier (BBB) that strictly
restricts drug penetration into the brain. Therefore, in central nervous
system drug development, the establishment of an in vitro human BBB model for use in studies estimating the in vivo human BBB permeability of drug candidates has long been awaited.
The current study developed and characterized a human immortalized
cell-based BBB triculture model, termed the “hiBBB”
model. To set up the hiBBB model, human immortalized BMEC (HBMEC/ci18)
were cocultured with human immortalized astrocytes (HASTR/ci35) and
brain pericytes (HBPC/ci37) in a transwell system. The trans-endothelial
electrical resistance of the hiBBB model was 134.4 ± 5.5 (Ω
× cm2), and the efflux ratios of rhodamine123 and
dantrolene were 1.72 ± 0.11 and 1.72 ± 0.45, respectively,
suggesting that the hiBBB model possesses essential cellular junction
and efflux transporter functions. In BBB permeability assays, the
mean value of the permeability coefficients (Pe) of BBB permeable compounds (propranolol, pyrilamine, memantine,
and diphenhydramine) was 960 × 10–6 cm/s, which
was clearly distinguishable from that of BBB nonpermeable compounds
(sodium fluorescein and Lucifer yellow, 18 × 10–6 cm/s). Collectively, this study successfully developed the hiBBB
model, which exhibits essential BBB functionality. Taking into consideration
the high availability of the immortalized cells used in the hiBBB
model, our results are expected to become an initial step toward the
establishment of a useful human BBB model to investigate drug penetration
into the human brain.