TY - JOUR
T1 - Large-Scale Production of LGR5-Positive Bipotential Human Liver Stem Cells
AU - Schneeberger, Kerstin
AU - Sánchez-Romero, Natalia
AU - Ye, Shicheng
AU - van Steenbeek, Frank G.
AU - Oosterhoff, Loes A.
AU - Pla Palacin, Iris
AU - Chen, Chen
AU - van Wolferen, Monique E.
AU - van Tienderen, Gilles
AU - Lieshout, Ruby
AU - Colemonts-Vroninks, Haaike
AU - Schene, Imre
AU - Hoekstra, Ruurdtje
AU - Verstegen, Monique M.A.
AU - van der Laan, Luc J.W.
AU - Penning, Louis C.
AU - Fuchs, Sabine A.
AU - Clevers, Hans
AU - De Kock, Joery
AU - Baptista, Pedro M.
AU - Spee, Bart
N1 - Publisher Copyright:
© 2019 The Authors. Hepatology published by Wiley Periodicals, Inc., on behalf of American Association for the Study of Liver Diseases.
PY - 2020/7/1
Y1 - 2020/7/1
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/85082971517
U2 - 10.1002/hep.31037
DO - 10.1002/hep.31037
M3 - Article
C2 - 31715015
AN - SCOPUS:85082971517
SN - 0270-9139
VL - 72
SP - 257
EP - 270
JO - Hepatology
JF - Hepatology
IS - 1
ER -