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Primary culture of choroidal epithelial cells: Characterization of an in vitro model of blood-CSF barrier

  • Cellular And Molecular Toxicology
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Summary

A primary rat choroidal epithelial cell culture system was developed to investigate mechanisms of heavy metal toxicity on the blood-cerebrospinal fluid (CSF) barrier. Epithelial cells were dissociated from choroidal tissue by pronase digestion and cultured in standard DMEM culture media supplemented with 10% fetal bovine serum and 10 ng epithelial growth factor per ml. The procedure yielded 2–5 × 104 cells from pooled plexuses of three to four rats, and a viability of 77–85%. The cultures displayed a dominant polygonal type of epithelial cells, with a population doubling time of 2–3 d. The cultures were of distinct choroidal epithelial origins. For example, immunocytochemical studies using monospecific rabbit anti-rat TTR polyclonal antibody revealed a strong positive stain of transthyretin (TTR), a thyroxine transport protein exclusively produced by the choroidal epithelia. Also, reverse-transcriptase polymerase chain reaction (PCR) confirmed the presence of specific TTR mRNA in the cultures. The cultures were further adapted to grow on a freely permeable membrane sandwiched between two culture chambers. The formation of an impermeable confluent monolayer occurred within 5 d after seeding and was verified by the presence of a steady electrical resistance across the membrane (80 ± 10 ohm per cm2). The epithelial barriers appeared to actively transport [125I]-thyroxine from the basal to apical chamber. These results suggest that this primary cell culture system possesses typical choroidal epithelial characteristics and appears to be a suitable model for in vitro mechanistic investigations of blood-CSF barrier.

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Authors and Affiliations

  1. Department of Pharmacology, College of Physicians and Surgeons, Columbia University, 10032, New York, New York

    Wei Zheng, QiuQu Zhao & Joseph H. Graziano

  2. Division of Environmental Health Sciences, Columbia University School of Public Health, 60 Haven Avenue, B1-112, 10032, New York, New York

    Wei Zheng & Joseph H. Graziano

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  1. Wei Zheng
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  2. QiuQu Zhao
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  3. Joseph H. Graziano
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Zheng, W., Zhao, Q. & Graziano, J.H. Primary culture of choroidal epithelial cells: Characterization of an in vitro model of blood-CSF barrier. In Vitro Cell.Dev.Biol.-Animal 34, 40–45 (1998). https://doi.org/10.1007/s11626-998-0051-8

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  • DOI: https://doi.org/10.1007/s11626-998-0051-8

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