Scale-up of hepatic progenitor cells from multitray stack to 2-D bioreactors

Promethera Biosciences (Mont-St-Guibert, BE) is developing cell therapies to treat several liver genetic metabolic diseases, such as the Crigler-Najjar syndrome. Human heterologous adult liver progenitors cells (HHALPCs) were initially cultivated in 2D standard cultivation devices. The present study is investigating the feasibility to cultivate HHALPCs in Xpansion bioreactors, with the following objectives: ➢ The process must be closed ➢ The growth rate and population-doubling level (i.e. the number of times the cells in the population has doubled) must be at least equivalent to the current process in multilayer trays ➢ The process must comply to the cGMP rules ➢ The cells must succeed the quality control (QC) test specifications at the end of cultivation, i.e. cells must remain undifferentiated and show the presence of HHAPLCs markers, while exhibiting the capacity to differentiate toward functional hepatocytes. Integrity® Xpansion™ multiplate bioreactors have been specifically designed to enable an easy transfer from existing multiple-tray-stack processes by offering the same cell growth environment on 2-D hydrophylized Polystyrene (PS) plates in a fully closed system. To make the bioreactors compact, the headspace between each plate has been reduced to a minimum (1.3 mm). Gas transfer is made through a semi-permeable silicone tubing mounted in the central column. Additionally, critical cell culture parameters such as pH and DO are controlled and the cell density is automatically monitored via a specific holographic microscope developed by Ovizio