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

In Vitro Cellular & Developmental Biology. Animal
W ZhengJ Graziano

Abstract

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 x 10(4) 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 ...Continue Reading

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Citations

Jun 6, 2013·Journal of Medicinal Food·Mohammad NabiuniJaleel A Miyan
Feb 15, 2011·Toxicology and Applied Pharmacology·Andrew D MonnotWei Zheng
Sep 11, 2007·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Lewis Zhichang ShiWei Zheng
Mar 31, 2015·PloS One·Basam Z Barkho, Edwin S Monuki
Jan 3, 2001·Microscopy Research and Technique·W Zheng
Aug 14, 2020·Brain Sciences·Shannon E MorganNingning Zhao
Aug 15, 2017·Brain Sciences·Robert P Lisak, Joyce A Benjamins

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