A Novel Pulse Damper for Endothelial Cell Flow Bioreactors.

Abstract

Peristaltic pumps (PP) are favored in flow bioreactors for their non-contact sterile design. But they produce pulsatile flow, which is consequential for the cultured cells. A novel pulse damper (PD) is reported for pulsatility elimination. ThePD designwas implemented to target static pressure pulsatility and flow rate (velocity) pulsatility from a PP. Damping effectiveness was tested in a macro-scale, closed-loop recirculating bioreactor mimicking the aortic arch at flow rates up to (4L/min). Time-resolved particle image velocimetry was used to characterize the velocity field. Endothelial cells (EC) were grown in the bioreactor, and subjected to continuous flow for 15min with or without PD. The PD was found to be nearly 90% effective at reducing pulsatility. The EC exposed to low PP flow without PD exhibited distress signaling in the form of increased ERK1/2 phosphorylation (2.5 folds) when compared to those exposed to the same flow with PD. At high pump flow without PD, the cells detached and did not survive, while they were perfectly healthy with PD. Flow pulsatility from PP causes EC distress at low flow and cell detachment at high flow. Elevated temporal shear stress gradient combined with elevated shear stress magnitude at high flow are believed to be the cause of cell detachment and death. The proposed PD design was effective at minimizing the hemodynamic stressors in the pump's output, demonstrably reducing cell distress. Adoption of the proposed PD design in flow bioreactors should improve experimental protocols.