Large‐scale Production of LGR5‐positive Bipotential Human Liver Stem Cells

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, Haaike Colemonts-Vroninks, Imre Schene, Ruurdtje Hoekstra, Monique M.A. Verstegen , Luc J.W. van der Laan , Louis C. Penning , Sabine A. Fuchs , Hans Clevers, Joery De Kock, Pedro M. Baptista Bart Spee

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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
Issue number1
Publication statusPublished - 1 Jul 2020

Bibliographical note

© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.

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