Clonal growth and serial propagation of rat esophageal epithelial cells. In Vitro 1983 May;19(5):403-15
Date
05/01/1983Pubmed ID
6345345DOI
10.1007/BF02619557Scopus ID
2-s2.0-0020980386 (requires institutional sign-in at Scopus site) 65 CitationsAbstract
The clonal growth and serial propagation of rat esophageal epithelial cells in low serum-containing medium has been achieved without feeder layers or conditioned medium. To date, a total of four lines have been developed and maintained for as many as 40 passages in culture. Growth of the cells was possible only after modifying the culture medium (PFMR-4) by reducing the calcium concentration from 1 to 0.1 mM, and by adding low levels of dialyzed fetal bovine serum and seven growth factors; i.e. epidermal growth factor, hydrocortisone, ethanolamine, phosphoethanolamine, insulin, transferrin, and cholera toxin. Cell lines have been developed from both explant outgrowths and enzyme dissociated esophagi. The epithelial nature of the cells was confirmed by electron microscopy and immunological methods. Clonal growth studies revealed that optimal cell growth occurred in medium containing 2.4% dialyzed fetal bovine serum and 0.1 mM calcium. Calcium levels of 0.3 mM or higher caused the cells to stratify and undergo terminal differentiation. Coating the culture dishes with collagen, or a combination of collagen, fibronectin, and bovine serum albumin, increased both the cell growth rate and the colony forming efficiency. The successful long term culture of rat esophageal epithelial cells permits their use as models in studies concerned with esophageal differentiation and carcinogenesis.
Author List
Babcock MS, Marino MR, Gunning WT 3rd, Stoner GDMESH terms used to index this publication - Major topics in bold
AnimalsCalcium
Cell Division
Cell Line
Clone Cells
Collagen
Culture Media
Endopeptidases
Epithelial Cells
Esophagus
Fetal Blood
Fibronectins
Growth Substances
Microscopy, Electron
Rats
