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 - Funding Information:
We thank Hans Vernooij for his help with the statistical analysis and Dr. Rupert Ecker from TissueGnostics GmbH, Austria, for his kind help on the quantification of the positive cell populations in the immunofluorescence presented in Fig. of this manuscript.
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
Y1 - 2020/7
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
SN - 0270-9139
VL - 72
SP - 257
EP - 270
JO - Hepatology (Baltimore, Md.)
JF - Hepatology (Baltimore, Md.)
IS - 1
ER -