Large-Scale Production of LGR5-Positive Bipotential Human Liver Stem Cells

Hepatology. 2020 Jul;72(1):257-270. doi: 10.1002/hep.31037. Epub 2020 Apr 8.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques*
  • Cell Differentiation
  • Cell Proliferation*
  • Cells, Cultured
  • Hepatocytes / cytology*
  • Humans
  • Liver Regeneration*
  • Liver Transplantation*
  • Organoids / cytology*
  • Receptors, G-Protein-Coupled / biosynthesis*
  • Stem Cells / metabolism*
  • Tissue Engineering*


  • LGR5 protein, human
  • Receptors, G-Protein-Coupled