Sheng-Li Huang, Xi-Jing He, Zong-Fang Li, Lu Yao, Wei Shi
{"title":"A novel primary culture method for rat choroidal epithelial cells.","authors":"Sheng-Li Huang, Xi-Jing He, Zong-Fang Li, Lu Yao, Wei Shi","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To establish a method for the culture of primary choroidal epithelial cells.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusion: </strong>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.</p>","PeriodicalId":520723,"journal":{"name":"Neurosciences (Riyadh, Saudi Arabia)","volume":" ","pages":"27-32"},"PeriodicalIF":0.0000,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Neurosciences (Riyadh, Saudi Arabia)","FirstCategoryId":"3","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Objective: 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.
Conclusion: 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.