Background: For biotechnological use of cells in tissue engineered applications, such as biohybrid renal devices, optimal culture conditions are required. Oxygen delivery is one of the most important cell determined system criterion for ex vivo applications. It is involved in the maintenance of highly oxygen-dependent renal tubular epithelial cells, affecting metabolic state, differentiation, and desired transport functions. The purpose of this study was to examine respiratory patterns such as basal oxygen consumption, solute transport-related oxygen demand, and oxygen concentration-dependent oxygen uptake of renal tubular epithelial cells in vitro. Methods: Respiratory patterns of highly purified human primary renal proximal (hPTC) and early distal tubular cells (hTALDC) were analyzed by perfusion respirometry. Spontaneous oxygen consumptions and maximum respirations after carbonyl cyanide m-chlorophenyl hydrazone (CCCP) uncoupling were measured. Respiration fractions contributing to basolateral Na+/K+-ATPase transport activities were assessed via ouabain inhibition and Na+-free medium. Furthermore, we determined oxygen uptake in dependency of oxygen concentration and morphology in various culture conditions (shaken, static). Results: Respiration of solely hPTC strongly depended on oxygen concentration in a Michaelis-Menten pattern at noncritical oxygen concentrations. Respiration of both cell types was significantly increased by CCCP, whereas average Na+/K+-ATPase-based oxygen uptake fractions differ significantly between the two cell types. Nevertheless, no significant differences were found in spontaneous respiration between hPTC and hTALDC. Conclusions: Our results clearly indicate that cell-specific oxygen consumption parameters have to be considered in the design of biotechnological devices intended to support kidney function by cell-supported renal replacement therapy. (C) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 27: 262-268, 2011