Defining conditions to promote the attachment of adult human colonic epithelial cells

Abstract

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.

Cell-cell contact and specific cytokines inhibit apoptosis of colonic epithelial cells: growth factors protect against c-myc-independent apoptosis

“…Limited success has been reported for short-term culture (2-4 days) (Buset et al, 1987) and, in our hands, using culture techniques developed for premalignant adenoma cells, normal colonic epithelial cells behave similarly (Paraskeva and Williams, 1992). We therefore hypothesize that specific survival factors must be required by normal colonic epithelium.…”

“…Buset et al (1987) have reported the short-term culture (2-4 days) of normal colonic epithelial cells, and our experience with such primary cultures is similar (Paraskeva and Williams, 1992). Isolated colonocytes only survive for a few days in culture.…”

Autofluorescence characterisation of isolated whole crypts and primary cultured human epithelial cells from normal, hyperplastic, and adenomatous colonic mucosa

“…29 30 There are also a few reports of short term normal cultures from adult colon tissues, but none in which the autofluorescence was studied. [31][32][33] In our present study, short term primary cell cultures of normal, hyperplastic, and dysplastic colonic epithelial cells obtained from biopsies were successfully established. Individual whole crypts were first isolated and imaged by confocal fluorescence microscopy (CFM).…”

Development of a three-dimensional model of lung cancer using cultured transformed lung cells

“…Another contemplated experiment will use molecular probes and proteomics that will clarify and characterize True cellular differentiation involves complex cellular interactions, [55][56][57] in which cell membrane junctions, extracellular matrices (e.g., basement membrane and ground substances) and soluble signals (endocrine, autocrine and paracrine) play important roles. [58][59][60][61] This process is also influenced by the spatial relationships (niche) of cells to each other. The responsiveness to exogenous mediators such as cytokines requires more than simply the presence of receptors.…”