Clonal growth and serial propagation of rat esophageal epithelial cells |
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Authors: | Merrill S Babcock Maria R Marino William T Gunning III Gary D Stoner |
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Institution: | (1) Department of Pathology, Medical College of Ohio, 3000 Arlington Avenue, 43614 Toledo, Ohio |
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Abstract: | Summary 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.
This investigation was supported by U.S. Public Health Service Grant CA 28950, awarded by the National Cancer Institute, Bethesda,
MD. |
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Keywords: | esophagus epithelial cells calcium growth factors substrates |
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