Volumetric Bioprinting of Organoids and Optically Tuned Hydrogels to Build Liver-Like Metabolic Biofactories

Paulina Nuñez Bernal; Manon Bouwmeester; Jorge Madrid-Wolff; Marc Falandt; Sammy Florczak; Nuria Ginés Rodriguez; Yang Li; Gabriel Größbacher; Roos-Anne Samsom; Monique van Wolferen; Luc van der Laan; Paul Delrot; Damien Loterie; Jos Malda; Christophe Moser; Bart Spee; Riccardo Levato

doi:10.1002/adma.202110054

Volumetric Bioprinting of Organoids and Optically Tuned Hydrogels to Build Liver-Like Metabolic Biofactories

Paulina Nuñez Bernal
, Manon Bouwmeester
, Jorge Madrid-Wolff
, Marc Falandt
, Sammy Florczak
, Nuria Ginés Rodriguez
, Yang Li
, Gabriel Größbacher
, Roos-Anne Samsom
, Monique van Wolferen
, Luc van der Laan
, Paul Delrot
, Damien Loterie
, Jos Malda
, Christophe Moser
, Bart Spee
, Riccardo Levato

Laboratory of Applied Photonics Devices
Utrecht University, University Medical Center Utrecht
Department of Virosciences, Erasmus Medical Center, Erasmus University, Rotterdam, The Netherlands.
EPFL Innovation Park

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Organ- and tissue-level biological functions are intimately linked to microscale cell–cell interactions and to the overarching tissue architecture. Together, biofabrication and organoid technologies offer the unique potential to engineer multi-scale living constructs, with cellular microenvironments formed by stem cell self-assembled structures embedded in customizable bioprinted geometries. This study introduces the volumetric bioprinting of complex organoid-laden constructs, which capture key functions of the human liver. Volumetric bioprinting via optical tomography shapes organoid-laden gelatin hydrogels into complex centimeter-scale 3D structures in under 20 s. Optically tuned bioresins enable refractive index matching of specific intracellular structures, countering the disruptive impact of cell-mediated light scattering on printing resolution. This layerless, nozzle-free technique poses no harmful mechanical stresses on organoids, resulting in superior viability and morphology preservation post-printing. Bioprinted organoids undergo hepatocytic differentiation showing albumin synthesis, liver-specific enzyme activity, and remarkably acquired native-like polarization. Organoids embedded within low stiffness gelatins (<2 kPa) are bioprinted into mathematically defined lattices with varying degrees of pore network tortuosity, and cultured under perfusion. These structures act as metabolic biofactories in which liver-specific ammonia detoxification can be enhanced by the architectural profile of the constructs. This technology opens up new possibilities for regenerative medicine and personalized drug testing.

Original language	English
Article number	2110054
Pages (from-to)	1-16
Journal	Advanced Materials
Volume	34
Issue number	15
Early online date	15 Feb 2022
DOIs	https://doi.org/10.1002/adma.202110054
Publication status	Published - 14 Apr 2022

Bibliographical note

Funding Information:
P.N.B. and M.B. contributed equally to this work. This project received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 949806, VOLUME‐BIO) and from the European's Union's Horizon 2020 research and innovation programme under grant agreement No 964497 (ENLIGHT). R.L and J.M acknowledge the funding from the ReumaNederland (LLP‐12, LLP22, and 19‐1‐207 MINIJOINT) and the Gravitation Program “Materials Driven Regeneration”, funded by the Netherlands Organization for Scientific Research (024.003.013). R.L. also acknowledges funding from the NWA‐Ideeëngenerator programme of the Netherlands Organization for Scientific Research (NWA.1228.192.105). M.B. and B.S. acknowledge funding from the research program Applied and Engineering Sciences with project number 15498, which is financed by the Netherlands Organization for Scientific Research. J.M.W. and C.M. acknowledge funding from the Swiss National Science Foundation: “Light based Volumetric printing in scattering resins” (n 200021_196971). L.J.W.vdL was supported by Medical Delta Program “Regenerative Medicine 4D”. The authors would like to thank D.J. Hall from Carbon and Neon for his support with scientific illustrations, V. Onink for his assistance with the particle tracking analysis, C. Spiegel for her help with editing, Dr. K. Schneeberger, and Dr. M. Pietribiasi for the fruitful discussions.

Funding Information:
P.N.B. and M.B. contributed equally to this work. This project received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 949806, VOLUME-BIO) and from the European's Union's Horizon 2020 research and innovation programme under grant agreement No 964497 (ENLIGHT). R.L and J.M acknowledge the funding from the ReumaNederland (LLP-12, LLP22, and 19-1-207 MINIJOINT) and the Gravitation Program ?Materials Driven Regeneration?, funded by the Netherlands Organization for Scientific Research (024.003.013). R.L. also acknowledges funding from the NWA-Idee?ngenerator programme of the Netherlands Organization for Scientific Research (NWA.1228.192.105). M.B. and B.S. acknowledge funding from the research program Applied and Engineering Sciences with project number 15498, which is financed by the Netherlands Organization for Scientific Research. J.M.W. and C.M. acknowledge funding from the Swiss National Science Foundation: ?Light based Volumetric printing in scattering resins? (n 200021_196971). L.J.W.vdL was supported by Medical Delta Program ?Regenerative Medicine 4D?. The authors would like to thank D.J. Hall from Carbon and Neon for his support with scientific illustrations, V. Onink for his assistance with the particle tracking analysis, C. Spiegel for her help with editing, Dr. K. Schneeberger, and Dr. M. Pietribiasi for the fruitful discussions.

Publisher Copyright:
© 2022 The Authors. Advanced Materials published by Wiley-VCH GmbH.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

biofabrication
bioresins
hydrogels
light-based 3D printing
volumetric additive manufacturing

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Advanced Materials - 2022 - Bernal - Volumetric Bioprinting of Organoids and Optically Tuned Hydrogels to Build Liver‐LikeFinal published version, 2.99 MBLicence: CC BY

Cite this

Bernal, P. N., Bouwmeester, M., Madrid-Wolff, J., Falandt, M., Florczak, S., Rodriguez, N. G., Li, Y., Größbacher, G., Samsom, R.-A., van Wolferen, M., van der Laan, L., Delrot, P., Loterie, D., Malda, J., Moser, C., Spee, B., & Levato, R. (2022). Volumetric Bioprinting of Organoids and Optically Tuned Hydrogels to Build Liver-Like Metabolic Biofactories. Advanced Materials, 34(15), 1-16. Article 2110054. https://doi.org/10.1002/adma.202110054

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title = "Volumetric Bioprinting of Organoids and Optically Tuned Hydrogels to Build Liver-Like Metabolic Biofactories",

abstract = "Organ- and tissue-level biological functions are intimately linked to microscale cell{\textendash}cell interactions and to the overarching tissue architecture. Together, biofabrication and organoid technologies offer the unique potential to engineer multi-scale living constructs, with cellular microenvironments formed by stem cell self-assembled structures embedded in customizable bioprinted geometries. This study introduces the volumetric bioprinting of complex organoid-laden constructs, which capture key functions of the human liver. Volumetric bioprinting via optical tomography shapes organoid-laden gelatin hydrogels into complex centimeter-scale 3D structures in under 20 s. Optically tuned bioresins enable refractive index matching of specific intracellular structures, countering the disruptive impact of cell-mediated light scattering on printing resolution. This layerless, nozzle-free technique poses no harmful mechanical stresses on organoids, resulting in superior viability and morphology preservation post-printing. Bioprinted organoids undergo hepatocytic differentiation showing albumin synthesis, liver-specific enzyme activity, and remarkably acquired native-like polarization. Organoids embedded within low stiffness gelatins (<2 kPa) are bioprinted into mathematically defined lattices with varying degrees of pore network tortuosity, and cultured under perfusion. These structures act as metabolic biofactories in which liver-specific ammonia detoxification can be enhanced by the architectural profile of the constructs. This technology opens up new possibilities for regenerative medicine and personalized drug testing.",

keywords = "biofabrication, bioresins, hydrogels, light-based 3D printing, volumetric additive manufacturing",

author = "Bernal, \{Paulina Nu{\~n}ez\} and Manon Bouwmeester and Jorge Madrid-Wolff and Marc Falandt and Sammy Florczak and Rodriguez, \{Nuria Gin{\'e}s\} and Yang Li and Gabriel Gr{\"o}{\ss}bacher and Roos-Anne Samsom and \{van Wolferen\}, Monique and \{van der Laan\}, Luc and Paul Delrot and Damien Loterie and Jos Malda and Christophe Moser and Bart Spee and Riccardo Levato",

note = "Funding Information: P.N.B. and M.B. contributed equally to this work. This project received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 949806, VOLUME‐BIO) and from the European's Union's Horizon 2020 research and innovation programme under grant agreement No 964497 (ENLIGHT). R.L and J.M acknowledge the funding from the ReumaNederland (LLP‐12, LLP22, and 19‐1‐207 MINIJOINT) and the Gravitation Program {\textquotedblleft}Materials Driven Regeneration{\textquotedblright}, funded by the Netherlands Organization for Scientific Research (024.003.013). R.L. also acknowledges funding from the NWA‐Idee{\"e}ngenerator programme of the Netherlands Organization for Scientific Research (NWA.1228.192.105). M.B. and B.S. acknowledge funding from the research program Applied and Engineering Sciences with project number 15498, which is financed by the Netherlands Organization for Scientific Research. J.M.W. and C.M. acknowledge funding from the Swiss National Science Foundation: {\textquotedblleft}Light based Volumetric printing in scattering resins{\textquotedblright} (n 200021\_196971). L.J.W.vdL was supported by Medical Delta Program {\textquotedblleft}Regenerative Medicine 4D{\textquotedblright}. The authors would like to thank D.J. Hall from Carbon and Neon for his support with scientific illustrations, V. Onink for his assistance with the particle tracking analysis, C. Spiegel for her help with editing, Dr. K. Schneeberger, and Dr. M. Pietribiasi for the fruitful discussions. Funding Information: P.N.B. and M.B. contributed equally to this work. This project received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 949806, VOLUME-BIO) and from the European's Union's Horizon 2020 research and innovation programme under grant agreement No 964497 (ENLIGHT). R.L and J.M acknowledge the funding from the ReumaNederland (LLP-12, LLP22, and 19-1-207 MINIJOINT) and the Gravitation Program ?Materials Driven Regeneration?, funded by the Netherlands Organization for Scientific Research (024.003.013). R.L. also acknowledges funding from the NWA-Idee?ngenerator programme of the Netherlands Organization for Scientific Research (NWA.1228.192.105). M.B. and B.S. acknowledge funding from the research program Applied and Engineering Sciences with project number 15498, which is financed by the Netherlands Organization for Scientific Research. J.M.W. and C.M. acknowledge funding from the Swiss National Science Foundation: ?Light based Volumetric printing in scattering resins? (n 200021\_196971). L.J.W.vdL was supported by Medical Delta Program ?Regenerative Medicine 4D?. The authors would like to thank D.J. Hall from Carbon and Neon for his support with scientific illustrations, V. Onink for his assistance with the particle tracking analysis, C. Spiegel for her help with editing, Dr. K. Schneeberger, and Dr. M. Pietribiasi for the fruitful discussions. Publisher Copyright: {\textcopyright} 2022 The Authors. Advanced Materials published by Wiley-VCH GmbH.",

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doi = "10.1002/adma.202110054",

language = "English",

volume = "34",

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Bernal, PN, Bouwmeester, M, Madrid-Wolff, J, Falandt, M, Florczak, S, Rodriguez, NG, Li, Y, Größbacher, G, Samsom, R-A, van Wolferen, M, van der Laan, L, Delrot, P, Loterie, D, Malda, J, Moser, C, Spee, B & Levato, R 2022, 'Volumetric Bioprinting of Organoids and Optically Tuned Hydrogels to Build Liver-Like Metabolic Biofactories', Advanced Materials, vol. 34, no. 15, 2110054, pp. 1-16. https://doi.org/10.1002/adma.202110054

TY - JOUR

T1 - Volumetric Bioprinting of Organoids and Optically Tuned Hydrogels to Build Liver-Like Metabolic Biofactories

AU - Bernal, Paulina Nuñez

AU - Bouwmeester, Manon

AU - Madrid-Wolff, Jorge

AU - Falandt, Marc

AU - Florczak, Sammy

AU - Rodriguez, Nuria Ginés

AU - Li, Yang

AU - Größbacher, Gabriel

AU - Samsom, Roos-Anne

AU - van Wolferen, Monique

AU - van der Laan, Luc

AU - Delrot, Paul

AU - Loterie, Damien

AU - Malda, Jos

AU - Moser, Christophe

AU - Spee, Bart

AU - Levato, Riccardo

N1 - Funding Information: P.N.B. and M.B. contributed equally to this work. This project received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 949806, VOLUME‐BIO) and from the European's Union's Horizon 2020 research and innovation programme under grant agreement No 964497 (ENLIGHT). R.L and J.M acknowledge the funding from the ReumaNederland (LLP‐12, LLP22, and 19‐1‐207 MINIJOINT) and the Gravitation Program “Materials Driven Regeneration”, funded by the Netherlands Organization for Scientific Research (024.003.013). R.L. also acknowledges funding from the NWA‐Ideeëngenerator programme of the Netherlands Organization for Scientific Research (NWA.1228.192.105). M.B. and B.S. acknowledge funding from the research program Applied and Engineering Sciences with project number 15498, which is financed by the Netherlands Organization for Scientific Research. J.M.W. and C.M. acknowledge funding from the Swiss National Science Foundation: “Light based Volumetric printing in scattering resins” (n 200021_196971). L.J.W.vdL was supported by Medical Delta Program “Regenerative Medicine 4D”. The authors would like to thank D.J. Hall from Carbon and Neon for his support with scientific illustrations, V. Onink for his assistance with the particle tracking analysis, C. Spiegel for her help with editing, Dr. K. Schneeberger, and Dr. M. Pietribiasi for the fruitful discussions. Funding Information: P.N.B. and M.B. contributed equally to this work. This project received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No. 949806, VOLUME-BIO) and from the European's Union's Horizon 2020 research and innovation programme under grant agreement No 964497 (ENLIGHT). R.L and J.M acknowledge the funding from the ReumaNederland (LLP-12, LLP22, and 19-1-207 MINIJOINT) and the Gravitation Program ?Materials Driven Regeneration?, funded by the Netherlands Organization for Scientific Research (024.003.013). R.L. also acknowledges funding from the NWA-Idee?ngenerator programme of the Netherlands Organization for Scientific Research (NWA.1228.192.105). M.B. and B.S. acknowledge funding from the research program Applied and Engineering Sciences with project number 15498, which is financed by the Netherlands Organization for Scientific Research. J.M.W. and C.M. acknowledge funding from the Swiss National Science Foundation: ?Light based Volumetric printing in scattering resins? (n 200021_196971). L.J.W.vdL was supported by Medical Delta Program ?Regenerative Medicine 4D?. The authors would like to thank D.J. Hall from Carbon and Neon for his support with scientific illustrations, V. Onink for his assistance with the particle tracking analysis, C. Spiegel for her help with editing, Dr. K. Schneeberger, and Dr. M. Pietribiasi for the fruitful discussions. Publisher Copyright: © 2022 The Authors. Advanced Materials published by Wiley-VCH GmbH.

PY - 2022/4/14

Y1 - 2022/4/14

N2 - Organ- and tissue-level biological functions are intimately linked to microscale cell–cell interactions and to the overarching tissue architecture. Together, biofabrication and organoid technologies offer the unique potential to engineer multi-scale living constructs, with cellular microenvironments formed by stem cell self-assembled structures embedded in customizable bioprinted geometries. This study introduces the volumetric bioprinting of complex organoid-laden constructs, which capture key functions of the human liver. Volumetric bioprinting via optical tomography shapes organoid-laden gelatin hydrogels into complex centimeter-scale 3D structures in under 20 s. Optically tuned bioresins enable refractive index matching of specific intracellular structures, countering the disruptive impact of cell-mediated light scattering on printing resolution. This layerless, nozzle-free technique poses no harmful mechanical stresses on organoids, resulting in superior viability and morphology preservation post-printing. Bioprinted organoids undergo hepatocytic differentiation showing albumin synthesis, liver-specific enzyme activity, and remarkably acquired native-like polarization. Organoids embedded within low stiffness gelatins (<2 kPa) are bioprinted into mathematically defined lattices with varying degrees of pore network tortuosity, and cultured under perfusion. These structures act as metabolic biofactories in which liver-specific ammonia detoxification can be enhanced by the architectural profile of the constructs. This technology opens up new possibilities for regenerative medicine and personalized drug testing.

AB - Organ- and tissue-level biological functions are intimately linked to microscale cell–cell interactions and to the overarching tissue architecture. Together, biofabrication and organoid technologies offer the unique potential to engineer multi-scale living constructs, with cellular microenvironments formed by stem cell self-assembled structures embedded in customizable bioprinted geometries. This study introduces the volumetric bioprinting of complex organoid-laden constructs, which capture key functions of the human liver. Volumetric bioprinting via optical tomography shapes organoid-laden gelatin hydrogels into complex centimeter-scale 3D structures in under 20 s. Optically tuned bioresins enable refractive index matching of specific intracellular structures, countering the disruptive impact of cell-mediated light scattering on printing resolution. This layerless, nozzle-free technique poses no harmful mechanical stresses on organoids, resulting in superior viability and morphology preservation post-printing. Bioprinted organoids undergo hepatocytic differentiation showing albumin synthesis, liver-specific enzyme activity, and remarkably acquired native-like polarization. Organoids embedded within low stiffness gelatins (<2 kPa) are bioprinted into mathematically defined lattices with varying degrees of pore network tortuosity, and cultured under perfusion. These structures act as metabolic biofactories in which liver-specific ammonia detoxification can be enhanced by the architectural profile of the constructs. This technology opens up new possibilities for regenerative medicine and personalized drug testing.

KW - biofabrication

KW - bioresins

KW - hydrogels

KW - light-based 3D printing

KW - volumetric additive manufacturing

UR - https://www.scopus.com/pages/publications/85125671084

U2 - 10.1002/adma.202110054

DO - 10.1002/adma.202110054

M3 - Article

C2 - 35166410

SN - 0935-9648

VL - 34

SP - 1

EP - 16

JO - Advanced Materials

JF - Advanced Materials

IS - 15

M1 - 2110054

ER -

Volumetric Bioprinting of Organoids and Optically Tuned Hydrogels to Build Liver-Like Metabolic Biofactories

Abstract

Bibliographical note

UN SDGs

Keywords

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