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Characterization of the Amino Acid Transport of New Immortalized Choroid Plexus Epithelial Cell Lines: A Novel In Vitro System for Investigating Transport Functions at the Blood-Cerebrospinal Fluid Barrier

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Abstract

Purpose. To establish and characterize a choroid plexus epithelial cell line (TR-CSFB) from a new type of transgenic rat harboring the temperature-sensitive simian virus 40 (ts SV 40) large T-antigen gene (Tg rat).

Methods. Choroid plexus epithelial cells were isolated from the Tg rat and cultured on a collagen-coated dish at 37°C during the first period of 3 days. Cells were subsequently cultured at 33°C to activate large T-antigen. At the third passage, cells were cloned by colony formation and isolated from other cells using a penicillin cup.

Results. Five immortalized cell lines of choroid plexus epithelial cells (TR-CSFB 1∼5) were obtained from two Tg rats. These cell lines had a polygonal cell morphology, expressed the typical choroid plexus epithelial cell marker, transthyretin, and possessed Na+, K+-ATPase on their apical side. TR-CSFBs cells expressed a large T-antigen and grew well at 33°C with a doubling-time of 35∼40 hr. [3H]-L-Proline uptake by TR-CSFB cells took place in an Na+-dependent, ouabain-sensitive, energy-dependent, and concentration-dependent manner. It was also inhibited by α-methylaminoisobutylic acid, suggesting that system A for amino acids operates in TR-CSFB cells. When [3H]-L-proline uptake was measured using the Transwell device, the L-proline uptake rate following application to the apical side was fivefold greater than that following application to the basal side. In addition, both Na+-dependent and Na+-independent L-glutamic acid (L-Glu) uptake processes were present in TR-CSFB cells.

Conclusions. Immortalized choroid plexus epithelial cell lines were successfully established from Tg rats and have the properties of choroid plexus epithelial cells, and amino acid transport activity was observed in vivo.

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Author notes

  1. Ken-ichi Hosoya

    Present address: Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama, 930-0194, Japan

Authors and Affiliations

  1. Department of Molecular Biopharmacy and Genetics, Graduate School of Pharmaceutical Sciences, Japan

    Takeo Kitazawa, Ken-ichi Hosoya, Masatomi Watanabe, Tadayuki Takashima, Sumio Ohtsuki, Hitomi Takanaga & Tetsuya Terasaki

  2. New Industry Creation Hatchery Center, Tohoku University, Aoba, Aramaki, Aoba-ku, Sendai, 980-8578, Japan

    Ken-ichi Hosoya, Sumio Ohtsuki, Hitomi Takanaga & Tetsuya Terasaki

  3. CREST of Japan Science and Technology Corporation (JST), Japan

    Ken-ichi Hosoya, Sumio Ohtsuki, Hitomi Takanaga, Masuo Obinata & Tetsuya Terasaki

  4. YS New Technology Institute Inc., 519 Shimoishibashi, Ishibashi-machi, Tochigi, 329-0512, Japan

    Masatsugu Ueda

  5. Department of Cell Biology, Institute of Development, Ageing and Cancer, Tohoku University, 4-1, Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan

    Nobuaki Yanai & Masuo Obinata

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Kitazawa, T., Hosoya, Ki., Watanabe, M. et al. Characterization of the Amino Acid Transport of New Immortalized Choroid Plexus Epithelial Cell Lines: A Novel In Vitro System for Investigating Transport Functions at the Blood-Cerebrospinal Fluid Barrier. Pharm Res 18, 16–22 (2001). https://doi.org/10.1023/A:1011014424212

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