A Novel Primary Culture Method for Rat Choroidal Epithelial Cells

Sheng-Li Huang, Xi-Jing He, Zong-Fang Li, Lu Yao, Wei Shi

ABSTRACT

Objectives: To establish a method for the culture of primary choroidal epithelial cells.

Methods: 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.

Results: 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.

Conclusions: 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.

Neurosciences Journal