Combination of culture on collagen gels and glucose starvation for cloning human colon cancer cells
| Autor: | Jean-Baptiste Rognoni, Maxime Lehmann, Jacques Fantini, Jacques Marvaldi, Chantal Rabenandrasana, Bernard Verrier |
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| Rok vydání: | 1991 |
| Předmět: |
Cell division
Cellular differentiation Clinical Biochemistry Biomedical Engineering Fluorescent Antibody Technique Bioengineering Cell Separation 03 medical and health sciences HT29 Cells 0302 clinical medicine Tumor Cells Cultured Humans 030304 developmental biology Cloning 0303 health sciences Confluency biology Chemistry Cell Differentiation Cell Biology Clone Cells Culture Media Cell biology Kinetics Microscopy Electron Mucus Glucose Cell culture 030220 oncology & carcinogenesis Colonic Neoplasms biology.protein Collagen Villin Cell Division Type I collagen Biotechnology |
| Zdroj: | Cytotechnology. 5:117-127 |
| ISSN: | 1573-0778 0920-9069 |
| DOI: | 10.1007/bf00365428 |
| Popis: | Glucose starvation has been widely used to select differentiated subpopulations from the heterogenous human colon cancer cell line HT29. We observed that the important cell loss elicited by culturing these cells in glucose-free medium could be limited when type I collagen gel was used as substratum instead of conventional plastic support. We took advantage of this property to develop a new protocol, which combined glucose starvation and culture on collagen gels, for cloning HT29 cells. Using this procedure we have isolated four clones that were characterized on the basis of morphological (optical and transmission electron microscopy), electrophysiological (determination of transepithelial electrical parameters) and biochemical (detection of villin, sucrase-isomaltase and carcinoembryonic antigen) criteria. These four clones expressed different patterns of enterocytic differentiation regarding to these criteria. These results confirmed the heterogeneity of the HT29 cell line. One of these clones, HT29-A7, which displayed numerous intercellular cysts that disappeared at confluency, appears as a complementary model in the study of epithelial biogenesis. |
| Databáze: | OpenAIRE |
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