Apical to basolateral surface area ratio and polarity of MDCK cells grown on different supports.
We have established conditions under which Madin-Darby canine kidney cells develop a well-polarized monolayer on polycarbonate filters and on transparent filters. These filters have biochemical and mechanical advantages over the nitrocellulose filters which have been widely used. Transepithelial resistance was established 10 h after plating and stabilized after 24 h. The distribution of protein antigens was followed by surface immunofluorescence and quantitated by a surface immunoassay that we developed. Uvomorulin was localized to the lateral membrane, with low amounts detectable on the basal membrane. The 58-kDa antigen was distributed over the entire basolateral domain, including cell processes extending into the filter pores. This distribution was confirmed by immunogold labeling of frozen sections. The 114-kDa antigen was found to be present at similar surface densities on both the apical and the basolateral domain. The support used for growth had profound effects on the cell morphology. A morphometric analysis of the plasma membrane of both strains of the cell line showed an increase in the number and size of the microvilli, and a smoother basal membrane as compared to published data on nitrocellulose filters. The apical to basolateral surface area ratio was therefore modified.
Butor
C
C
European Molecular Biology Laboratory, Heidelberg, Germany.
Davoust
J
J
D000818:Animals; D015415:Biomarkers; D002455:Cell Division; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure*; D046508:Culture Techniques / Q000379:methods; D004285:Dogs; D004847:Epithelial Cells; D004848:Epithelium / Q000502:physiology / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D007668:Kidney; D007700:Kinetics; D008565:Membrane Proteins / Q000032:analysis; D008854:Microscopy, Electron; D008871:Microvilli / Q000648:ultrastructure; D011108:Polymers
D015415:Biomarkers; D008565:Membrane Proteins; D011108:Polymers