• Kerstin Schneeberger
  • Natalia Sánchez‐Romero
  • Shicheng Ye
  • Frank G van Steenbeek
  • Loes A Oosterhoff
  • Iris Pla Palacin
  • Chen Chen
  • Monique E. van Wolferen
  • Gilles van Tienderen
  • Ruby Lieshout
  • Imre Schene
  • Ruurdtje Hoekstra
  • Monique M.A. Verstegen
  • Luc J.W. van der Laan
  • Louis C. Penning
  • Sabine A. Fuchs
  • Hans Clevers
  • Pedro M. Baptista
  • Bart Spee

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.

Original languageEnglish
Pages (from-to)257-270
Number of pages14
JournalHepatology
Volume72
Issue number1
DOIs
Publication statusPublished - 1 Jul 2020

ID: 49234809