Self-Organogenesis from 2D Micropatterns to 3D Biomimetic Biliary Trees
Abstract
:- Biliary ducts have been precluded from previous liver bioengineering studies.
- Bile ducts were generated from cholangiocytes self-organization on micropatterns.
- Co-culture with endothelial cells allowed the formation of millimeter long biliary networks with interconnected lumens.
- This is the first model of intrahepatic biliary ducts of defined geometry.
1. Introduction
2. Results
2.1. Bile Duct Trees Formed by Self-Organization from Micropatterns
2.2. Study of the Luminogenesis of Bile Duct Structures
2.3. Characterization of Bile Duct Lumens
2.4. Bile Duct Structures Retain Apico-Basal Polarity with Preserved Cholangiocyte Phenotype
2.5. Trees Can Be Detached and Remain Functional
2.6. Study of the Self-Organization Process
3. Discussion
4. Material and Methods
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
3D | three dimensional |
2D | bidimensional |
ECM | Extracellular matrix |
NRCs or N | Normal Rat Cholangiocytes |
HUVECs or H | Human Umbilical Vein Endothelial Cells |
FDA | Fluorescein Diacetate |
References
- Acun, A.; Oganesyan, R.; Uygun, B.E. Liver Bioengineering: Promise, Pitfalls, and Hurdles to Overcome. Curr. Transplant. Rep. 2019, 6, 119–126. [Google Scholar] [CrossRef] [PubMed]
- Vyas, D.; Baptista, P.M.; Brovold, M.; Moran, E.; Gaston, B.; Booth, C.; Samuel, M.; Atala, A.; Soker, S. Self-assembled liver organoids recapitulate hepatobiliary organogenesis in vitro. Hepatology 2018, 67, 750–761. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Verstegen, M.M.A.; Roos, F.J.M.; Burka, K.; Gehart, H.; Jager, M.; de Wolf, M.; Bijvelds, M.J.C.; de Jonge, H.R.; Ardisasmita, A.I.; van Huizen, N.A.; et al. Human extrahepatic and intrahepatic cholangiocyte organoids show region-specific differentiation potential and model cystic fibrosis-related bile duct disease. Sci. Rep. 2020, 10, 1–16. [Google Scholar] [CrossRef]
- Tanimizu, N.; Ichinohe, N.; Sasaki, Y.; Itoh, T.; Sudo, R.; Yamaguchi, T.; Katsuda, T.; Ninomiya, T.; Tokino, T.; Ochiya, T.; et al. Generation of functional liver organoids on combining hepatocytes and cholangiocytes with hepatobiliary connections ex vivo. Nat. Commun. 2021, 12, 1–12. [Google Scholar] [CrossRef] [PubMed]
- Huang, Y.; Sakai, Y.; Hara, T.; Katsuda, T.; Ochiya, T.; Gu, W.L.; Eguchi, S. Bioengineering of a CLiP-derived tubular biliary-duct-like structure for bile transport in vitro. Biotechnol. Bioeng. 2021, 118, 2572–2584. [Google Scholar] [CrossRef] [PubMed]
- Roos, F.J.M.; Verstegen, M.M.A.; Albarinos, L.M.; Roest, H.P.; Poley, J.-W.; Tetteroo, G.W.M.; Ijzermans, J.N.M.; van der Laan, L.J.W. Human Bile Contains Cholangiocyte Organoid-Initiating Cells Which Expand as Functional Cholangiocytes in Non-canonical Wnt Stimulating Conditions. Front. Cell Dev. Biol. 2020, 8, 630492. [Google Scholar] [CrossRef]
- Justin, A.W.; Saeb-Parsy, K.; Markaki, A.E.; Vallier, L.; Sampaziotis, F. Advances in the generation of bioengineered bile ducts. Biochim. Biophys. Acta (BBA) Mol. Basis Dis. 2018, 1864, 1532–1538. [Google Scholar] [CrossRef]
- Buisson, E.M.; Jeong, J.; Kim, H.J.; Choi, D. Regenerative Medicine of the Bile Duct: Beyond the Myth. Int. J. Stem Cells 2019, 12, 183–194. [Google Scholar] [CrossRef]
- Li, X.; Wen, L.; Liu, J.; Wang, X. Three-dimensional Printing-Driving Liver Therapies. Curr. Med. Chem. 2021, 28, 1. [Google Scholar] [CrossRef]
- Jung, D.J.; Byeon, J.H.; Jeong, G.S. Flow enhances phenotypic and maturation of adult rat liver organoids. Biofabrication 2020, 12, 045035. [Google Scholar] [CrossRef]
- Smith, Q.; Chen, C.; Bhatia, S. Directing Cholangiocyte Morphogenesis in Natural Biomaterial Scaffolds. bioRxiv 2021. [Google Scholar] [CrossRef]
- Chen, C.; Jochems, P.G.M.; Salz, L.; Schneeberger, K.; Penning, L.C.; Van De Graaf, S.F.J.; Beuers, U.; Clevers, H.; Geijsen, N.; Masereeuw, R.; et al. Bioengineered bile ducts recapitulate key cholangiocyte functions. Biofabrication 2018, 10, 034103. [Google Scholar] [CrossRef]
- Tysoe, O.C.; Justin, A.W.; Brevini, T.; Chen, S.E.; Mahbubani, K.T.; Frank, A.K.; Zedira, H.; Melum, E.; Saeb-Parsy, K.; Markaki, A.E.; et al. Isolation and propagation of primary human cholangiocyte organoids for the generation of bioengineered biliary tissue. Nat. Protoc. 2019, 14, 1884–1925. [Google Scholar] [CrossRef]
- Du, Y.; Khandekar, G.; Llewellyn, J.; Polacheck, W.; Chen, C.S.; Wells, R.G. A Bile Duct-on-a-Chip with Organ-Level Functions. Hepatology 2020, 71, 1350–1363. [Google Scholar] [CrossRef]
- Lewis, P.L.; Su, J.; Yan, M.; Meng, F.; Glaser, S.S.; Alpini, G.D.; Green, R.M.; Sosa-Pineda, B.; Shah, R.N. Complex bile duct network formation within liver decellularized extracellular matrix hydrogels. Sci. Rep. 2018, 8, 12220. [Google Scholar] [CrossRef] [PubMed]
- Lewis, P.L.; Yan, M.; Su, J.; Shah, R.N. Directing the growth and alignment of biliary epithelium within extracellular matrix hydrogels. Acta Biomater. 2019, 85, 84–93. [Google Scholar] [CrossRef] [PubMed]
- Théry, M. Micropatterning as a tool to decipher cell morphogenesis and functions. J. Cell Sci. 2010, 123, 4201–4213. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hauser, P.V.; Nishikawa, M.; Kimura, H.; Fujii, T.; Yanagawa, N. Controlled tubulogenesis from dispersed ureteric bud-derived cells using a micropatterned gel. J. Tissue Eng. Regen. Med. 2016, 10, 762–771. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tsang, K.M.; Annabi, N.; Ercole, F.; Zhou, K.; Karst, D.; Li, F.; Haynes, J.M.; Evans, R.A.; Thissen, H.; Khademhosseini, A.; et al. Facile One-step Micropatterning Using Photodegradable Methacrylated Gelatin Hydrogels for Improved Cardiomyocyte Organization and Alignment. Adv. Funct. Mater. 2015, 25, 977–986. [Google Scholar] [CrossRef] [Green Version]
- Bosch-Fortea, M.; Rodriguez-Fraticelli, A.E.; Herranz, G.; Hachimi, M.; Barea, M.D.; Young, J.; Ladoux, B.; Martin-Belmonte, F. Martin-Belmonte Micropattern-based platform as a physiologically relevant model to study epithelial morphogenesis and nephrotoxicity. Biomaterials 2019, 218, 119339. [Google Scholar] [CrossRef]
- Jeon, O.; Alsberg, E. Spatial micropatterning of growth factors in three-dimensional hydrogels for location-specific regulation of cellular behaviors. Small 2018, 14, e1800579. [Google Scholar] [CrossRef]
- Zouani, O.F.; Lei, Y.; Durrieu, M.-C. Pericytes, Stem-Cell-Like Cells, but not Mesenchymal Stem Cells are Recruited to Support Microvascular Tube Stabilization. Small 2013, 9, 3070–3075. [Google Scholar] [CrossRef]
- Bouzhir, L.; Gontran, E.; Loarca, L.; Collado-Hilly, M.; Dupuis-Williams, P. Generation and Quantitative Characterization of Functional and Polarized Biliary Epithelial Cysts. J. Vis. Exp. 2020, 159, e61404. [Google Scholar] [CrossRef]
- Daniel, E.; Cleaver, O. Vascularizing organogenesis: Lessons from developmental biology and implications for regenerative medicine. Curr. Top. Dev. Biol. 2019, 132, 177–220. [Google Scholar] [PubMed]
- Tanimizu, N.; Kaneko, K.; Itoh, T.; Ichinohe, N.; Ishii, M.; Mizuguchi, T.; Hirata, K.; Miyajima, A.; Mitaka, T. Intrahepatic bile ducts are developed through formation of homogeneous continuous luminal network and its dynamic rearrangement in mice. Hepatology 2016, 64, 175–188. [Google Scholar] [CrossRef] [PubMed]
- Ober, E.A.; Lemaigre, F.P. Development of the liver: Insights into organ and tissue morphogenesis. J. Hepatol. 2018, 68, 1049–1062. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tabibian, J.H.; Masyuk, A.I.; Masyuk, T.V.; O’Hara, S.P.; LaRusso, N.F. Physiology of Cholangiocytes. Compr. Physiol. 2013, 3, 541–565. [Google Scholar]
- Gigliozzi, A.; Fraioli, F.; Sundaram, P.; Lee, J.; Mennone, A.; Alvaro, D.; Boyer, J.L. Molecular identification and functional characterization of Mdr1a in rat cholangiocytes. Gastroenterol. 2000, 119, 1113–1122. [Google Scholar] [CrossRef]
- Vroman, B.; LaRusso, N.F. Development and characterization of polarized primary cultures of rat intrahepatic bile duct epithelial cells. Lab. Investig. 1996, 74, 303–313. [Google Scholar]
- Iruela-Arispe, M.; Beitel, G. Tubulogenesis. Development 2013, 140, 2851–2855. [Google Scholar] [CrossRef] [Green Version]
- Rodriguez-Fraticelli, A.; Auzan, M.; Alonso, M.; Bornens, M.; Martin-Belmonte, F. Cell confinement controls centrosome positioning and lumen initiation during epithelial morphogenesis. J. Cell Biol. 2012, 198, 1011–1023. [Google Scholar] [CrossRef] [Green Version]
- Enemchukwu, N.O.; Cruz-Acuña, R.; Bongiorno, T.; Johnson, C.; García, J.R.; Sulchek, T.; García, A.J. Synthetic matrices reveal contributions of ECM biophysical and biochemical properties to epithelial morphogenesis. J. Cell Biol. 2016, 212, 113–124. [Google Scholar] [CrossRef] [Green Version]
- Funfak, A.; Bouzhir, L.; Gontran, E.; Minier, N.; Dupuis-Williams, P.; Gobaa, S. Biophysical Control of Bile Duct Epithelial Morphogenesis in Natural and Synthetic Scaffolds. Front. Bioeng. Biotechnol. 2019, 7, 417. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Si-Tayeb, K.; Lemaigre, F.; Duncan, S.A. Organogenesis and Development of the Liver. Dev. Cell 2010, 18, 175–189. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Baptista, P.M.; Moran, E.; Vyas, D.; Shupe, T.; Soker, S. Liver regeneration and bioengineering: The role of liver extra-cellular matrix and human stem/progenitor cells. In Regenerative Medicine Applications in Organ Transplantation; Academic Press: Cambridge, MA, USA, 2014; pp. 391–400. [Google Scholar]
- Fabris, L.; Fiorotto, R.; Spirli, C.; Cadamuro, M.; Mariotti, V.; Perugorria, M.J.; Banales, J.M.; Strazzabosco, M. Pathobiology of inherited biliary diseases: A roadmap to understand acquired liver diseases. Nat. Rev. Gastroenterol. Hepatol. 2019, 16, 497–511. [Google Scholar] [CrossRef] [PubMed]
- Azioune, A.; Carpi, N.; Tseng, Q.; Théry, M.; Piel, M. Protein micropatterns: A direct printing protocol using deep UVs. Methods Cell Biol. 2010, 97, 133–146. [Google Scholar]
- Schindelin, J.; Arganda-Carreras, I.; Frise, E.; Kaynig, V.; Longair, M.; Pietzsch, T.; Preibisch, S.; Rueden, C.; Saalfeld, S.; Schmid, B.; et al. Fiji: An open-source platform for biological-image analysis. Nat. Methods 2012, 9, 676–682. [Google Scholar] [CrossRef] [Green Version]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Gontran, E.; Loarca, L.; El Kassis, C.; Bouzhir, L.; Ayollo, D.; Mazari-Arrighi, E.; Fuchs, A.; Dupuis-Williams, P. Self-Organogenesis from 2D Micropatterns to 3D Biomimetic Biliary Trees. Bioengineering 2021, 8, 112. https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering8080112
Gontran E, Loarca L, El Kassis C, Bouzhir L, Ayollo D, Mazari-Arrighi E, Fuchs A, Dupuis-Williams P. Self-Organogenesis from 2D Micropatterns to 3D Biomimetic Biliary Trees. Bioengineering. 2021; 8(8):112. https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering8080112
Chicago/Turabian StyleGontran, Emilie, Lorena Loarca, Cyrille El Kassis, Latifa Bouzhir, Dmitry Ayollo, Elsa Mazari-Arrighi, Alexandra Fuchs, and Pascale Dupuis-Williams. 2021. "Self-Organogenesis from 2D Micropatterns to 3D Biomimetic Biliary Trees" Bioengineering 8, no. 8: 112. https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering8080112