Evaluation of Na+ active transport and morphological changes for bioartificial renal tubule cell device using Madin-Darby canine kidney cells.

Abstract

The function of current hemodialysis as an artificial kidney is insufficient because of the lack of reabsorptive function. In this study, we intend to develop a bioartificial renal tubule cell device (RTD) 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 the culture period on their ability to transport Na+ actively using Na+/K+ATPase (NKA). The results were (1) active Na+ transport of the cells averaged 24.8 mM/m(2) x 24 h during the initial 2 weeks after confluence and then decreased to about 4.2 mM/m(2) x 24 h during the next 2 weeks; (2) NKA localized on the basal-lateral sides of the cells during the initial 2 weeks, whereas it also localized on the apical side of the cells during the next 2 weeks; (3) long-term culture resulted in an increased number of upheaving cell mass, increased fatty droplets in the cells, and necrosis; and (4) scanning electron microscopy showed fewer microvilli 3-four weeks after confluence. It is concluded that the culture period is critical for developing RTD using cultured renal tubular epithelial cells.