Oxygen consumption characteristics in 3D constructs depend on cell density

Oxygen is not only crucial for cell survival but also a determinant for cell fate and function. However, the supply of oxygen and other nutrients as well as the removal of toxic waste products often limit cell viability in 3-dimensional (3D) engineered tissues. The aim of this study was to determine the oxygen consumption characteristics of 3D constructs as a function of their cell density. The oxygen concentration was measured at the base of hepatocyte laden constructs and a tightly controlled experimental and analytical framework was used to reduce the system geometry to a single coordinate and enable the precise identification of initial and boundary conditions. Then dynamic process modeling was used to fit the measured oxygen vs. time profiles to a reaction and diffusion model. We show that oxygen consumption rates are well-described by Michaelis-Menten kinetics. However, the reaction parameters are not literature constants but depend on the cell density. Moreover, the average cellular oxygen consumption rate (or OCR) also varies with density. We discuss why the OCR of cells is often misinterpreted and erroneously reported, particularly in the case of 3D tissues and scaffolds.