Cell culture distribution in a three-dimensional porous scaffold in perfusion bioreactor

Abstract

MotivationIt has been widely demonstrated that perfusion bioreactors improve in vitro cell culture on three-dimensional (3D) matrices. GoalWe aimed to determine the local cell distribution, based on the pore diameter, within a 3D scaffold during cell growth in a perfusion bioreactor. Materials and methodsScaffolds under perfusion were collected at 0, 4, and 7 days of culture; scaffold morphology and cell distribution within the construct were evaluated by imaging analysis. A mathematical model representing the cell growth heterogeneity in 3D perfusion bioreactor was developed; a specific rate equation for glucose was experimentally determined. Model accuracy was assessed comparing the cell proliferation predictions with experimental data. ResultsCell proliferation was successfully determined by coupling cell imaging with an ad hoc algorithm. The total number of cells in the scaffold, the number of cells for a single cross section, and the local cell distribution based on the scaffold pore size, were determined. Good agreement between computations and experiments was observed. ConclusionsA new methodology was developed to determine the cell heterogeneity within a 3D scaffold under perfusion. The proposed approach provides a helpful tool for 3D dynamic cell culture optimization for biological studies and tissue engineering applications.