Perfusion Circuit Concepts for Hollow-Fiber Bioreactors Used as in Vitro Cell Production Systems or Ex Vivo Bioartificial Organs

Abstract

For the development and implementation of primary human cell- and stem cell-based applications in regenerative medicine, large amounts of cells with well-defined characteristics are needed. Such cell quantities can be obtained with the use of hollow fiber-based bioreactors. While the use of such bioreactors generally requires a perfusion circuit, the configuration and complexity of such circuits is still in debate. We evaluated various circuit configurations to investigate potential perfusate volume shifts in the arterial and venous sides of the perfusion circuit, as well as in the feed and waste lines. Volume shifts with changes in flow conditions were measured with graduated bubble traps in the circuit, and perfusion pressures were measured at three points in the circuits. The results of this study demonstrate that the bioreactor perfusion circuit configuration has an effect on system pressures and volume shifts in the circuit. During operation, spikes in post-bioreactor pressures caused detrimental, potentially dangerous volume shifts in the feed and waste lines for configurations that lacked feed pumps and/or waste line check valves. Our results indicate that a more complex tubing circuit adds to safety of operation and avoids technical challenges associated with the use of large-scale hollow fiber bioreactors (e.g., for extracorporeal liver support or erythrocyte production from hematopoietic stem cells), including volume shifts and the need for a large reservoir. Finally, to ensure safe use of bioreactors, measuring pre-, intra-, and post-bioreactor pressures, and pump operation control is also advisable, which suggests the use of specifically developed bioreactor perfusion devices.