Defining conditions to promote the attachment of adult human colonic epithelial cells.
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.
D001485:Basement Membrane / Q000502:physiology; D001706:Biopsy; D002448:Cell Adhesion; D002455:Cell Division; D002478:Cells, Cultured; D003094:Collagen; D003106:Colon / Q000166:cytology*; D003470:Culture Media; D004847:Epithelial Cells; D005353:Fibronectins; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010100:Oxygen / Q000494:pharmacology; D012710:Serum Albumin, Bovine
D003470:Culture Media; D005353:Fibronectins; D012710:Serum Albumin, Bovine; D003094:Collagen; D010100:Oxygen