Madin Darby Canine Kidney (MDCK) Cells: Early & Late Passage

ABSTRACT: The paracellular passage of ions in epithelial systems is dictated by the repertoire of tight junction (TJ) proteins, the number and architecture of apicolateral TJ strands, and the interaction of TJ proteins with the cytoskeleton. To investigate the relative contribution of these factors and to explore their regulation, we examined a passage number dependent phenotypic change in Madin Darby Canine Kidney (MDCK) cells iteratively passaged at low density in which dilution potentials of the monolayer abruptly decreased but transepithelial resistance (TER) progressively increased. Here, we report that the phenotypic transition involved a decrease surface expression of peanut agglutinin (PNA), a decrease in steady state levels of claudin-1 and a slight decrease in E-cadherin expression in the context of an apparently claudin-2-negative system. Late passage cells grew in more densely packed monolayers and exhibited characteristics of a competent, functional cultured monolayer including inducible vectorial ion transport in response to norepinephrine (NE) and sphere formation in three dimensional laminin gel culture. Microarray analysis of gene expression levels in early and late passage cells revealed an increase in transcript abundance for SGK and GLUT-3, and a reduction Id-3. The steady state protein expression levels of GLUT-3, however, were found to be equivalent. These findings suggest that the expression of claudin-2 in MDCK cells could be contingent on signaling via lateral cell contacts and that claudin-1, in the context of sustained claudin-8 expression, could function as a negative regulator of overall TER. 3 biological replicates of early and late passage were run on Affymetrix chips. the replicates were averaged then compared.

ORGANISM(S): Canis lupus familiaris

SUBMITTER: Virginie Aris 

PROVIDER: E-GEOD-14837 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress