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Title: CONFOCAL LASER SCANNING MICROSCOPY IMAGING OF AN IN VITRO MODEL OF THE BOVINE BLOOD-MILK BARRIER

Author
item SMITS, E - U GHENT, BELGIUM
item BURVENICH, C - U CHENT, BELGIUM
item VAN OOSTVELDT, P - U GHENT, BELGIUM
item Guidry, Albert
item MASSART-LEEN, A - U GHENT, BELGIUM

Submitted to: Insect Neurochemistry and Neurophysiology Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 4/1/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Confocal affords increased resolution and three-dimensional optical sectioning. It preserves tissue architecture and allows for multi labeling. Three-dimensional imaging was performed on an in vitro model of the blood-milk barrier of the bovine mammary gland. The model consisted of a monolayer of bovine primary mammary artery endothelial cells on an matrix of bovine fibroblasts and calf-skin collagen type I, which in-turn was layered on a monolayer of bovine primary mammary ductal epithelial cells. Immunochemistry was used to identify the monolayers. The blue fluorescent dye DAPI was used to stain cell nuclei. The endothelial cells specifically stained for von Willebrand factor inside Weibel-Palade bodies. Fibroblasts were identified with antibody to vimentin, which is largely found in cells of mesenchymal origin. Cytokeratin filaments unique to mammary epithelial cells appeared as interconnected bundles in the epithelial cell cytoplasm. Protein synthesis was used to determine functional differentiation of the epithelial cells. Protein synthesis was detected using immunocytochemical staining with FITC-labeled antibodies to specific milk proteins. The epithelial cells stained positive for a-casein, the major protein of bovine milk, indicating fully differentiated cells. Alpha-lactalbumin was present in the apical region of the epithelial cells, indicating cell differentiation and the polar nature of the epithelial cells. Heterogenous differentiation of the epithelial cells, was indicative of the variation among cells in primary cell cultures. In conclusion, confocal laser scanning microscopy provides a useful tool in the study of in vitro mammary gland biology.