Preparation of a bioartificial kidney using tubular epithelial cells, and an evaluation of Na+ active transport and morphological changes

We intend to develop a bioartificial kidney using tubular epithelial cells and artificial membranes, and to evaluate the reabsorptive function of the confluent layers. Madin-Darby canine kidney (MDCK) cells were cultured on a nucleopore polycarbonate membrane for up to 4 weeks after confluence to examine the influence of culture period on their properties, such as the localization of Na^sup +^/K^sup +^-ATPase and active Na^sup +^ transport. The results were as follows. Ouabain-sensitive Na^sup +^ active transport declined at 3 to 4 weeks after confluence in each matrix. The localization of Na^sup +^/K^sup +^-ATPase indicated depolarization in the cell membrane 3 to 4 weeks after confluence. Prolongation of the culture period increased the formation of an upheaving cell mass after the formation of the confluent monolayer. Scanning electron microscopy revealed fewer microvilli and more flat cells after 3 to 4 weeks of confluency. We conclude that the decline of Na^sup +^ active transport in the MDCK cells was due to both the formation of multilayers and a decline of cell function throughout the long period of culture following the formation of the confluent monolayers. Further study for selection of membrane material, the extracellular matrix, and species of cells should be continued.[PUBLICATION ABSTRACT]