Extracellular matrix and mouse mammary cell function: Comparison of substrata in culture
| Autor: | C. J. Wilde, K. A. K. Hendry, Walter L. Hurley, D. R. Blatchford |
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| Rok vydání: | 1994 |
| Předmět: |
Cell Count
Epithelium Extracellular matrix Mice chemistry.chemical_compound Mammary Glands Animal Cell Adhesion medicine Protein biosynthesis Animals Secretion Cells Cultured chemistry.chemical_classification biology Lactoferrin Transferrin Caseins Proteins Cell Differentiation Epithelial Cells Cell Biology General Medicine Culture Media Extracellular Matrix EGTA Secretory protein chemistry Biochemistry Protein Biosynthesis Microscopy Electron Scanning biology.protein Collagenase Female Developmental Biology medicine.drug |
| Zdroj: | In Vitro Cellular & Developmental Biology - Animal. 30:529-538 |
| ISSN: | 1543-706X 1071-2690 |
| DOI: | 10.1007/bf02631326 |
| Popis: | Cultured mammary cells depend on interaction with a substratum for functional differentiation, even in the presence of lactogenic hormones. Protein synthesis and secretion by mouse mammary epithelial cells on floating collagen gels and (EHS) matrix were compared. Cells were prepared by collagenase digestion of tissue from mid-pregnant mice. Protein synthesis was consistently greater in cells attached to EHS matrix, and was associated with proportionately higher rates of protein secretion into culture medium. Cells on EHS secreted protein into a luminal space formed within multicellular alveolus-like structures. Luminal secreted protein, extracted by EGTA treatment of cells in situ, constituted up to 40% of total secreted radiolabeled protein for cells on EHS matrix. The EGTA extract contained a higher proportion of casein and lactoferrin, whereas transferrin was predominantly in the medium. This indicated that cells on EHS matrix had become polarized and were secreting proteins vectorially. In contrast, EGTA treatment of cells on floating collagen gels released virtually no radiolabeled protein, showing that mammosphere formation was a property of cells on EHS. These biochemical observations were supported by ultrastructural evidence. In EHS cultures, the proportion of secreted protein in the luminal fraction, but not the distribution of secreted proteins, changed with time. This suggests that there may be leakage out of the lumen, or intraluminal degradation of protein after secretion. Nevertheless, the results suggest that cellular organization into mammospheres on EHS matrix promotes synthetic and secretory activity. This system provides a useful model for investigation of the regulation of milk secretion. |
| Databáze: | OpenAIRE |
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