A novel primary culture method for rat choroidal epithelial cells
Huang, S.-L.; He, X.-J.; Li, Z.-F.; Yao, L.; Shi, W.
Neurosciences 18(1): 27-32
2013
ISSN/ISBN: 1319-6138 PMID: 23291794 Document Number: 667798
To establish a method for the culture of primary choroidal epithelial cells. This descriptive experimental study was carried out in Xi`an Jiaotong University, Xi`an, China from September 2009 to August 2012. Choroidal epithelial cells were isolated from the choroid plexus tissues of the lateral ventricles from neonatal rats (n=36). The tissues were dissociated into small cell aggregates by a mechanical method, and cultured on plastic culture dishes containing Dulbecco`s modified Eagle`s medium with 10% fetal bovine serum and 10 ng/ml epidermal growth factor at 37 degrees C in an incubator with 5% humidified carbon dioxide. The cultured cells were examined by phase contrast microscope, electron microscopy, and immunocytochemistry. The cells showed typical morphologic characteristics of epithelial phenotypes with a cobblestone appearance in monolayer 7-9 days post-seeding. The electron microscopy spotted typical choroidal epithelial cells with microvilli on the cytomembrane, organelles in the cytoplasm, and tight junctions welding 2 adjacent cells. They were positive against anti-transthyretin immunostaining. This culture technique, which does not require complex equipment and operation skills, might be a simple and efficient method for obtaining choroidal epithelial cells in sufficient number and purity from mixed primary cultures of rat tissue.