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

doi:10.1002/hep.31037

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 BaptistaBart Spee

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

92 Citations (Scopus)

Abstract

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. Here, we established a new 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 two weeks, compared to 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 upregulated compared to static organoid cultures, and cytochrome p450 activity reached levels equivalent to hepatocytes. CONCLUSION: 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 language	English
Journal	Hepatology
DOIs	https://doi.org/10.1002/hep.31037
Publication status	E-pub ahead of print - 12 Nov 2019

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10.1002/hep.31037

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Schneeberger, K., Sánchez-Romero, N., Ye, S., van Steenbeek, F. G., Oosterhoff, L. A., Pla Palacin, I., Chen, C., van Wolferen, M. E., van Tienderen, G., Lieshout, R., Colemonts-Vroninks, H., Schene, I., Hoekstra, R., Verstegen, M. M. A., van der Laan, L. J. W., Penning, L. C., Fuchs, S. A., Clevers, H., De Kock, J., ... Spee, B. (2019). Large-scale Production of LGR5-positive Bipotential Human Liver Stem Cells. Hepatology. Advance online publication. https://doi.org/10.1002/hep.31037

@article{f776bc982e9c4cd28b08548756fcb3d9,

title = "Large-scale Production of LGR5-positive Bipotential Human Liver Stem Cells",

abstract = "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. Here, we established a new 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 two weeks, compared to 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 upregulated compared to static organoid cultures, and cytochrome p450 activity reached levels equivalent to hepatocytes. CONCLUSION: 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.",

author = "Kerstin Schneeberger and Natalia S{\'a}nchez-Romero and Shicheng Ye and {van Steenbeek}, {Frank G} and Oosterhoff, {Loes A} and {Pla Palacin}, Iris and Chen Chen and {van Wolferen}, {Monique E} and {van Tienderen}, Gilles and Ruby Lieshout and Haaike Colemonts-Vroninks and Imre Schene and Ruurdtje Hoekstra and Verstegen, {Monique M A} and {van der Laan}, {Luc J W} and Penning, {Louis C} and Fuchs, {Sabine A} and Hans Clevers and {De Kock}, Joery and Baptista, {Pedro M} and Bart Spee",

note = "{\textcopyright} 2019 The Authors. Hepatology published by Wiley Periodicals, Inc. on behalf of American Association for the Study of Liver Diseases.",

year = "2019",

month = nov,

day = "12",

doi = "10.1002/hep.31037",

language = "English",

journal = "Hepatology",

issn = "0270-9139",

publisher = "American Association for the Study of Liver Diseases (AASLD)",

}

Schneeberger, K, Sánchez-Romero, N, Ye, S, van Steenbeek, FG, Oosterhoff, LA, Pla Palacin, I, Chen, C, van Wolferen, ME, van Tienderen, G, Lieshout, R, Colemonts-Vroninks, H, Schene, I, Hoekstra, R, Verstegen, MMA, van der Laan, LJW, Penning, LC, Fuchs, SA, Clevers, H, De Kock, J, Baptista, PM & Spee, B 2019, 'Large-scale Production of LGR5-positive Bipotential Human Liver Stem Cells', Hepatology. https://doi.org/10.1002/hep.31037

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

PY - 2019/11/12

Y1 - 2019/11/12

N2 - 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. Here, we established a new 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 two weeks, compared to 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 upregulated compared to static organoid cultures, and cytochrome p450 activity reached levels equivalent to hepatocytes. CONCLUSION: 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 - 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. Here, we established a new 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 two weeks, compared to 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 upregulated compared to static organoid cultures, and cytochrome p450 activity reached levels equivalent to hepatocytes. CONCLUSION: 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.

U2 - 10.1002/hep.31037

DO - 10.1002/hep.31037

M3 - Article

C2 - 31715015

SN - 0270-9139

JO - Hepatology

JF - Hepatology

ER -

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

Abstract

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