Influence of several extracellular matrix components in primary cultures of bovine mammary epithelial cells.
Mammary epithelial cells, obtained from lactating cows, were cultured onto inserts coated with several components of extracellular matrix. The influence of these components upon the maintenance of differentiation has been determinated. Every day, alpha S1-casein secretion was measured by radioimmunoassay (RIA) in apical and basal compartments. Reorganization of functional tight junctions was evaluated by measurement of transepithelial electrical resistance (TER). On EHS matrix, cells underwent alveolar structures and never established TER. alpha S1-casein secretion strongly fluctuated with the day of culture. When plated onto fibronectin, cells reorganized a typical pavement and established TER. Nevertheless, TER and casein secretion highly fluctuated. On laminin-coated inserts, a few cells bound to the substratum, dedifferentiated, and proliferated to confluency within 9 days. TER progressively increased to a stable level after 15 days. Casein was not recovered after 6 days. Cells on type I collagen-coated inserts reorganized an epithelial pavement within 2 days and quickly established a stable TER. They secreted apically high levels of casein during 2 weeks. As cells maintained their biochemical differentiation, the culture on type I collagen-coated inserts seems an efficient model for primary culture of bovine mammary epithelial cells and allows studies of polarized alpha S1-casein secretion.
Delabarre
S
S
Laboratoire de Sciences Animales, INRA-ENSAIA, Vandoeuvre, France. sezoot@ensaia.u-nancy.fr
Claudon
C
C
Laurent
F
F
D000818:Animals; D002364:Caseins / Q000378:metabolism; D002417:Cattle; D002454:Cell Differentiation; D002478:Cells, Cultured; D004594:Electrophysiology; D004848:Epithelium; D005109:Extracellular Matrix / Q000502:physiology*; D005260:Female; D006801:Humans; D008321:Mammary Glands, Animal / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D019108:Tight Junctions
D002364:Caseins