Defining conditions to promote the attachment of adult human colonic epithelial cells |
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Authors: | Michel Buset Sidney Winawer Eileen Friedman |
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Affiliation: | (1) Laboratory of Gastrointestinal Cancer Research and Gastroenterology Service, Department of Medicine, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 5614, 10021 New York, New York;(2) Present address: Hospital Erasme, Bruxelles, Belgium |
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Abstract: | Summary An improved method for the attachment and growth of normal human colonic epithelial cells from minute 1 to 3-mm3 biopsies has been developed. This yields four times as many cultured cells per biopsy than older methods, with a success rate of 97% in a series of 29 biopsies. Fetal bovine serum was eliminated from the medium, the medium pH was decreased to 6.7, the oxygen tension in the incubator was decreased to 3%, and the NCTC 168 medium was supplemented with ethanolamine, phosphoethanolamine, hydrocortisone, ascorbic acid, transferrin, glutamine, insulin, epidermal growth factor, pentagastrin, and deoxycholic acid. The best substrate for cell attachment was a mixture of ungelled collagen I and bovine serum albumin. This substrate was better than the identical mixture with fibronectin added, fibronectin alone, a thin gelatin film, collagen IV with or without fibronectin, and basement membrane preparations from four different cell lines. |
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Keywords: | normal colonic epithelial cell culture |
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