Investigation of the hepatic development in the coculture of hiPSCs-derived LSECs and HLCs in a fluidic microenvironment

Interactions between the different liver cell types are critical to the maintenance or induction of their function in vitro. In this work, human-induced Pluripotent Stem Cells (hiPSCs)-derived Liver Sinusoidal Endothelial Cells (LSECs) and Hepatocytes-Like Cells (HLCs) were cultured and matured in a...

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Main Authors: Mathieu Danoy, Yannick Tauran, Stephane Poulain, Rachid Jellali, Johanna Bruce, Marjorie Leduc, Morgane Le Gall, Yuta Koui, Hiroshi Arakawa, Francoise Gilard, Bertrand Gakiere, Yukio Kato, Charles Plessy, Taketomo Kido, Atsushi Miyajima, Yasuyuki Sakai, Eric Leclerc
Format: Article
Language:English
Published: AIP Publishing LLC 2021-06-01
Series:APL Bioengineering
Online Access:http://dx.doi.org/10.1063/5.0041227
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author Mathieu Danoy
Yannick Tauran
Stephane Poulain
Rachid Jellali
Johanna Bruce
Marjorie Leduc
Morgane Le Gall
Yuta Koui
Hiroshi Arakawa
Francoise Gilard
Bertrand Gakiere
Yukio Kato
Charles Plessy
Taketomo Kido
Atsushi Miyajima
Yasuyuki Sakai
Eric Leclerc
spellingShingle Mathieu Danoy
Yannick Tauran
Stephane Poulain
Rachid Jellali
Johanna Bruce
Marjorie Leduc
Morgane Le Gall
Yuta Koui
Hiroshi Arakawa
Francoise Gilard
Bertrand Gakiere
Yukio Kato
Charles Plessy
Taketomo Kido
Atsushi Miyajima
Yasuyuki Sakai
Eric Leclerc
Investigation of the hepatic development in the coculture of hiPSCs-derived LSECs and HLCs in a fluidic microenvironment
APL Bioengineering
author_facet Mathieu Danoy
Yannick Tauran
Stephane Poulain
Rachid Jellali
Johanna Bruce
Marjorie Leduc
Morgane Le Gall
Yuta Koui
Hiroshi Arakawa
Francoise Gilard
Bertrand Gakiere
Yukio Kato
Charles Plessy
Taketomo Kido
Atsushi Miyajima
Yasuyuki Sakai
Eric Leclerc
author_sort Mathieu Danoy
title Investigation of the hepatic development in the coculture of hiPSCs-derived LSECs and HLCs in a fluidic microenvironment
title_short Investigation of the hepatic development in the coculture of hiPSCs-derived LSECs and HLCs in a fluidic microenvironment
title_full Investigation of the hepatic development in the coculture of hiPSCs-derived LSECs and HLCs in a fluidic microenvironment
title_fullStr Investigation of the hepatic development in the coculture of hiPSCs-derived LSECs and HLCs in a fluidic microenvironment
title_full_unstemmed Investigation of the hepatic development in the coculture of hiPSCs-derived LSECs and HLCs in a fluidic microenvironment
title_sort investigation of the hepatic development in the coculture of hipscs-derived lsecs and hlcs in a fluidic microenvironment
publisher AIP Publishing LLC
series APL Bioengineering
issn 2473-2877
publishDate 2021-06-01
description Interactions between the different liver cell types are critical to the maintenance or induction of their function in vitro. In this work, human-induced Pluripotent Stem Cells (hiPSCs)-derived Liver Sinusoidal Endothelial Cells (LSECs) and Hepatocytes-Like Cells (HLCs) were cultured and matured in a microfluidic environment. Both cell populations were differentiated in Petri dishes, detached, and inoculated in microfluidic biochips. In cocultures of both cell types, the tissue has exhibited a higher production of albumin (3.19 vs 5.31 μg/mL/106 cells in monocultures and cocultures) as well as a higher inducibility CYP450 over monocultures of HLCs. Tubular-like structures composed of LSECs and positive for the endothelial marker PECAM1, as well as a tissue more largely expressing Stabilin-2 were detected in cocultures only. In contrast, monocultures exhibited no network and less specific endothelial markers. The transcriptomic analysis did not reveal a marked difference between the profiles of both culture conditions. Nevertheless, the analysis allowed us to highlight different upstream regulators in cocultures (SP1, EBF1, and GATA3) and monocultures (PML, MECP2, and NRF1). In cocultures, the multi-omics dataset after 14 days of maturation in biochips has shown the activation of signaling related to hepatic maturation, angiogenesis, and tissue repair. In this condition, inflammatory signaling was also found to be reduced when compared to monocultures as illustrated by the activation of NFKB and by the detection of several cytokines involved in tissue injury in the latter. Finally, the extracted biological processes were discussed regarding the future development of a new generation of human in vitro hepatic models.
url http://dx.doi.org/10.1063/5.0041227
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spelling doaj-e70436d4d10a4e2393e6907ba57b1f842021-07-08T13:17:23ZengAIP Publishing LLCAPL Bioengineering2473-28772021-06-0152026104026104-1310.1063/5.0041227Investigation of the hepatic development in the coculture of hiPSCs-derived LSECs and HLCs in a fluidic microenvironmentMathieu Danoy0Yannick Tauran1Stephane Poulain2Rachid Jellali3Johanna Bruce4Marjorie Leduc5Morgane Le Gall6Yuta Koui7Hiroshi Arakawa8Francoise Gilard9Bertrand Gakiere10Yukio Kato11Charles Plessy12Taketomo Kido13Atsushi Miyajima14Yasuyuki Sakai15Eric Leclerc16 CNRS UMI 2820, Laboratory for Integrated Micro Mechatronic Systems, Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan CNRS UMI 2820, Laboratory for Integrated Micro Mechatronic Systems, Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan RIKEN Center for Life Science Technologies, Division of Genomic Technologies, 1-7-22, Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan Université de Technologie de Compiègne, CNRS, Biomechanics and Bioengineering, Centre de recherche Royallieu, CS 60319, 60203 Compiègne Cedex, Compiegne, France Plateforme protéomique 3P5, Université de Paris, Institut Cochin, INSERM, CNRS, F-75014 Paris, France Plateforme protéomique 3P5, Université de Paris, Institut Cochin, INSERM, CNRS, F-75014 Paris, France Plateforme protéomique 3P5, Université de Paris, Institut Cochin, INSERM, CNRS, F-75014 Paris, France Laboratory of Stem Cell Therapy, Institute for Quantitative Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan Laboratory of Molecular Pharmacokinetics, Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa City, Ishikawa 920-1192, Japan Institute of Plant Sciences Paris-Saclay (IPS2), UMR 9213/UMR1403, CNRS, INRA, Université Paris-Sud, Université d'Evry, Université Paris-Diderot, Sorbonne Paris-Cité, Saclay Plant Sciences, Bâtiment 630, 91405 Orsay, France Institute of Plant Sciences Paris-Saclay (IPS2), UMR 9213/UMR1403, CNRS, INRA, Université Paris-Sud, Université d'Evry, Université Paris-Diderot, Sorbonne Paris-Cité, Saclay Plant Sciences, Bâtiment 630, 91405 Orsay, France Laboratory of Molecular Pharmacokinetics, Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kakuma-machi, Kanazawa City, Ishikawa 920-1192, Japan RIKEN Center for Life Science Technologies, Division of Genomic Technologies, 1-7-22, Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan Laboratory of Stem Cell Therapy, Institute for Quantitative Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan Laboratory of Stem Cell Therapy, Institute for Quantitative Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan Department of Chemical System Engineering, graduate school of Engineering, University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan CNRS UMI 2820, Laboratory for Integrated Micro Mechatronic Systems, Institute of Industrial Science, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, JapanInteractions between the different liver cell types are critical to the maintenance or induction of their function in vitro. In this work, human-induced Pluripotent Stem Cells (hiPSCs)-derived Liver Sinusoidal Endothelial Cells (LSECs) and Hepatocytes-Like Cells (HLCs) were cultured and matured in a microfluidic environment. Both cell populations were differentiated in Petri dishes, detached, and inoculated in microfluidic biochips. In cocultures of both cell types, the tissue has exhibited a higher production of albumin (3.19 vs 5.31 μg/mL/106 cells in monocultures and cocultures) as well as a higher inducibility CYP450 over monocultures of HLCs. Tubular-like structures composed of LSECs and positive for the endothelial marker PECAM1, as well as a tissue more largely expressing Stabilin-2 were detected in cocultures only. In contrast, monocultures exhibited no network and less specific endothelial markers. The transcriptomic analysis did not reveal a marked difference between the profiles of both culture conditions. Nevertheless, the analysis allowed us to highlight different upstream regulators in cocultures (SP1, EBF1, and GATA3) and monocultures (PML, MECP2, and NRF1). In cocultures, the multi-omics dataset after 14 days of maturation in biochips has shown the activation of signaling related to hepatic maturation, angiogenesis, and tissue repair. In this condition, inflammatory signaling was also found to be reduced when compared to monocultures as illustrated by the activation of NFKB and by the detection of several cytokines involved in tissue injury in the latter. Finally, the extracted biological processes were discussed regarding the future development of a new generation of human in vitro hepatic models.http://dx.doi.org/10.1063/5.0041227