Oxygen consumption rate of cells in 3D culture: The use of experiment and simulation to measure kinetic parameters and optimise culture conditions

Abstract

Understanding the basal O₂ and nutrient requirements of cells is paramount when culturing cells in 3D tissue models. Any scaffold design will need to take such parameters into consideration, especially as the addition of cells introduces gradients of consumption of such molecules from the surface to the core of scaffolds. We have cultured two cell types in 3D native collagen type I scaffolds, and measured the O₂ tension at specific locations within the scaffold. By changing the density of cells, we have established O₂ consumption gradients within these scaffolds and using mathematical modeling have derived rates of consumption for O₂. For human dermal fibroblasts the average rate constant was 1.19 × 10⁻¹⁷ mol cell⁻¹ s⁻¹, and for human bone marrow derived stromal cells the average rate constant was 7.91 × 10⁻¹⁸ mol cell⁻¹ s⁻¹. These values are lower than previously published rates for similar cells cultured in 2D, but the values established in this current study are more representative of rates of consumption measured in vivo. These values will dictate 3D culture parameters, including maximum cell-seeding density and maximum size of the constructs, for long-term viability of tissue models.