Background and Aims: The gap between patients on transplant waiting lists and available donor organs is steadily increasing. Human organoids derived from leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5)–positive adult stem cells represent an exciting new cell source for liver regeneration; however, culturing large numbers of organoids with current protocols is tedious and the level of hepatic differentiation is limited. Approach and Results: Here, we established a method for the expansion of large quantities of human liver organoids in spinner flasks. Due to improved oxygenation in the spinner flasks, organoids rapidly proliferated and reached an average 40-fold cell expansion after 2 weeks, compared with 6-fold expansion in static cultures. The organoids repopulated decellularized liver discs and formed liver-like tissue. After differentiation in spinner flasks, mature hepatocyte markers were highly up-regulated compared with static organoid cultures, and cytochrome p450 activity reached levels equivalent to hepatocytes. Conclusions: We established a highly efficient method for culturing large numbers of LGR5-positive stem cells in the form of organoids, which paves the way for the application of organoids for tissue engineering and liver transplantation.

dx.doi.org/10.1002/hep.31037, hdl.handle.net/1765/126143
Hepatology
Department of Surgery

Schneeberger, K, Sánchez-Romero, N. (Natalia), Ye, S. (Shicheng), van Steenbeek, F.G, Oosterhoff, L.A. (Loes A.), Pla Palacin, I. (Iris), … Spee, B. (2019). Large-Scale Production of LGR5-Positive Bipotential Human Liver Stem Cells. Hepatology. doi:10.1002/hep.31037