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Large‐Scale Production of LGR5‐Positive Bipotential Human Liver Stem Cells
Author(s) -
Schneeberger Kerstin,
SánchezRomero Natalia,
Ye Shicheng,
Steenbeek Frank G.,
Oosterhoff Loes A.,
Pla Palacin Iris,
Chen Chen,
Wolferen Monique E.,
Tienderen Gilles,
Lieshout Ruby,
ColemontsVroninks Haaike,
Schene Imre,
Hoekstra Ruurdtje,
Verstegen Monique M.A.,
Laan Luc J.W.,
Penning Louis C.,
Fuchs Sabine A.,
Clevers Hans,
De Kock Joery,
Baptista Pedro M.,
Spee Bart
Publication year - 2020
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.31037
Subject(s) - organoid , lgr5 , laboratory flask , stem cell , microbiology and biotechnology , biology , transplantation , liver cytology , hepatocyte , decellularization , chemistry , medicine , cancer stem cell , biochemistry , liver metabolism , in vitro , extracellular matrix
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.