Isolation, characterization, and long-term culture of fetal bovine tracheal epithelial cells
| Autor: | Marian L. Miller, Brenda L. Schumann, Terence E. Cody, George D. Leikauf |
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| Rok vydání: | 1988 |
| Předmět: |
Plating efficiency
Clinical Biochemistry Connective tissue Cell Separation Plant Science Biology Endopeptidases Freezing Dispase Keratin medicine Animals Trypsin Cells Cultured chemistry.chemical_classification Microvilli Tight junction Cell Membrane Epithelial Cells Cell Biology Anatomy Epithelium Culture Media Extracellular Matrix Trypsinization Cell biology Trachea Dithiothreitol Microscopy Electron Intercellular Junctions medicine.anatomical_structure chemistry Cell culture Karyotyping Keratins Cattle Collagen Glass Tissue Preservation Cell Division Biotechnology Developmental Biology |
| Zdroj: | In Vitro Cellular & Developmental Biology. 24:211-216 |
| ISSN: | 1475-2689 0883-8364 |
| DOI: | 10.1007/bf02623549 |
| Popis: | Epithelial cells were isolated from fetal bovine trachea by exposing and stripping the mucosal epithelium from the adjacent connective tissue. The tissue was minced and enzymically dissociated in Ca-Mg-free medium containing dispase and dithiothreitol. The stripping procedure and selective trypsinization produced epithelial cell cultures free of fibroblasts. Seeded on plastic, the plating efficiency was 21.5% with a doubling time of 24 h. Dome formation, evidence of occluding junctions and active ion transport characteristic of epithelial cells, was common. Growth of the cells on glass, collagen, and Engelbreth-Holm-Swarm (EHS) substrate demonstrated a striking difference in morphology. Cells grown on EHS presented a more distinctly three-dimensional growth pattern and many more microvilli when compared to cells grown on glass or collagen. The cells retained their epithelioid characteristics through more than 30 passages as shown by the presence of distinct apical and basolateral membranes, tight junctions, and positive keratin staining. |
| Databáze: | OpenAIRE |
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