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A well plate–based multiplexed platform for incorporation of organoids into an organ-on-a-chip system with a perfusable vasculature

Nature Protocols volume 16, pages 2158–2189 (2021)Cite this article

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Abstract

Owing to their high spatiotemporal precision and adaptability to different host cells, organ-on-a-chip systems are showing great promise in drug discovery, developmental biology studies and disease modeling. However, many current micro-engineered biomimetic systems are limited in technological application because of culture media mixing that does not allow direct incorporation of techniques from stem cell biology, such as organoids. Here, we describe a detailed alternative method to cultivate millimeter-scale functional vascularized tissues on a biofabricated platform, termed ‘integrated vasculature for assessing dynamic events’, that enables facile incorporation of organoid technology. Utilizing the 3D stamping technique with a synthetic polymeric elastomer, a scaffold termed ‘AngioTube’ is generated with a central microchannel that has the mechanical stability to support a perfusable vascular system and the self-assembly of various parenchymal tissues. We demonstrate an increase in user familiarity and content analysis by situating the scaffold on a footprint of a 96-well plate. Uniquely, the platform can be used for facile connection of two or more tissue compartments in series through a common vasculature. Built-in micropores enable the studies of cell invasion involved in both angiogenesis and metastasis. We describe how this protocol can be applied to create both vascularized cardiac and hepatic tissues, metastatic breast cancer tissue and personalized pancreatic cancer tissue through incorporation of patient-derived organoids. Platform assembly to populating the scaffold with cells of interest into perfusable functional vascularized tissue will require 12–14 d and an additional 4 d if pre-polymer and master molds are needed.

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Fig. 1: Dimensions of the InVADE platform and AngioTube bioscaffolds.
Fig. 2: Polymer synthesis setup and assessment of the 1,2,4 pre-polymer.
Fig. 3: Microfabrication of master molds for the AngioTube scaffold and base plate using photoresist and a soft lithography technique.
Fig. 4: Microfabrication of the AngioTube scaffold through the 3D stamping technique.
Fig. 5: Assembly of the InVADE platform.
Fig. 6: Vascularization and parenchymal tissue seeding around the AngioTube scaffold on an InVADE platform.
Fig. 7: Engineering functional vascularized 3D tissues on the InVADE platform.
Fig. 8: Engineering integrated organ-on-a-chip microdevices with a duo-organ InVADE platform for cancer metastasis study.

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Data availability

All data presented in this paper are available from the original references, the source data files supplied with this publication and the corresponding author. Source data are provided with this paper.

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Acknowledgements

This work was funded by the Canadian Institutes of Health Research (CIHR) Operating Grants (MOP-126027, MOP-137107 and MOP-142382), Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant (RGPIN 326982-10), NSERC-CIHR Collaborative Health Research Grant (CHRP 493737-16), CIHR Foundation Grant (FDN-167274) and National Institutes of Health Grant 2R01 HL076485. M.R. was supported by a Canada Research Chair and Killam Fellowship, B.F.L.L. and R.X.Z.L. were supported by a NSERC Postgraduate Fellowship and L.D.H. was supported by a CIHR Vanier Scholarship.

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Authors and Affiliations

  1. Institute of Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada

    Benjamin Fook Lun Lai, Rick Xing Ze Lu, Locke Davenport Huyer, Erika Yan Wang, Qinghua Wu & Milica Radisic

  2. Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada

    Locke Davenport Huyer, Joshua Yazbeck & Milica Radisic

  3. Faculty of Chemistry, Materials, and Bioengineering, Kansai University, Osaka, Japan

    Sachiro Kakinoki

  4. Organization for Research and Development of Innovative Science and Technology, Kansai University, Osaka, Japan

    Sachiro Kakinoki

  5. Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada

    Boyang Zhang

  6. Toronto General Research Institute, University Health Network, Toronto, Ontario, Canada

    Milica Radisic

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Contributions

B.F.L.L., B.Z. and M.R. designed the research. B.F.L.L. and L.D.H. performed the research. B.F.L.L. analyzed the data. B.F.L.L., S.K., L.D.H. and R.X.Z.L prepared the figures. B.F.L.L., R.X.Z.L. and J.Y. prepared the supplementary videos. B.F.L.L., L.D.H. and M.R. wrote and edited the manuscript. B.F.L.L. and E.Y.W. prepared the fluorescent images. Q.W. provided the induced-pluripotent stem cell–derived cardiomyocytes for cardiac tissue engineering.

Corresponding author

Correspondence to Milica Radisic.

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Competing interests

M.R. and B.Z. are among the co-founders of TARA Biosystems, and they hold equity in this company. The AngioTube bioscaffold is licensed to TARA Biosystems. The remaining authors declare no competing interests.

Additional information

Peer review information Nature Protocols thanks Christopher C. W. Hughes, Noo Li Jeon and Ibrahim Tarik Ozbolat for their contribution to the peer review of this work.

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Related links

Key references using this protocol

Lai, B. F. L. et al. Adv. Funct. Mat. 27, 1703524 (2017): https://doi.org/10.1002/adfm.201703524

Lai, B. F. L. et al. Adv. Funct. Mat. 30, 2000545 (2020): https://doi.org/10.1002/adfm.202000545

Lu, R. X. Z. et al. Adv. Mat. Tech. (2020): https://doi.org/10.1002/admt.202000726

Supplementary information

Supplementary Information

Supplementary Figs. 1–5 and Supplementary Text 1.

Supplementary Video 1

Preparation of AngioTube scaffolds before the cleanroom.

Supplementary Video 2

Assembling AngioTube scaffolds with 3D stamping.

Supplementary Video 3

Hot-embossing of InVADE base plates.

Supplementary Video 4

Assembling InVADE plates.

Supplementary Video 5

Endothelialization of the AngioTube scaffold lumen on the InVADE platform.

Supplementary Video 6

Tracking cancer metastasis on a duo-organ model of the InVADE platform.

Supplementary Video 7

Perfusion of 1-µm FITC beads in the InVADE platform.

Supplementary Data 1

Photomasks for soft lithography microfabrication.

Source data

Source Data Fig. 2

Excel data for the stress-strain curve of the 2:3 version of the 1,2,4 polymer.

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Lai, B.F.L., Lu, R.X.Z., Davenport Huyer, L. et al. A well plate–based multiplexed platform for incorporation of organoids into an organ-on-a-chip system with a perfusable vasculature. Nat Protoc 16, 2158–2189 (2021). https://doi.org/10.1038/s41596-020-00490-1

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