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Reconfigurable open microfluidics for studying the spatiotemporal dynamics of paracrine signalling

Nature Biomedical Engineering volume 3, pages 830–841 (2019)Cite this article

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Abstract

The study of intercellular signalling networks requires organotypic microscale systems that facilitate the culture, conditioning and manipulation of cells. Here, we describe a reconfigurable microfluidic cell-culture system that facilitates the assembly of three-dimensional tissue models by stacking layers that contain preconditioned microenvironments. By using principles of open and suspended microfluidics, the Stacks system is easily assembled or disassembled to provide spatial and temporal manoeuvrability in two-dimensional and three-dimensional assays of multiple cell types, enabling the modelling of sequential paracrine-signalling events, such as tumour-cell-mediated differentiation of macrophages and macrophage-facilitated angiogenesis. We used Stacks to recapitulate the in vivo observation that different prostate cancer tissues polarize macrophages with distinct gene-expression profiles of pro-inflammatory and anti-inflammatory cytokines. Stacks also enabled us to show that these two types of macrophages signal distinctly to endothelial cells, leading to blood vessels with different morphologies. Our proof-of-concept experiments exemplify how Stacks can efficiently model multicellular interactions and highlight the importance of spatiotemporal specificity in intercellular signalling.

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Fig. 1: Schematic and concept of Stacks, a reconfigurable multilayer suspended microfluidic system.
Fig. 2: Principles of suspended microfluidics enable layers to be stacked without bonding and without leaking outside the channels.
Fig. 3: Reconfigurable Stacks system enables addition and removal of macrophages at different stages to study chronological effects in cell signalling.
Fig. 4: A sequential Stacks assay was performed to achieve organotypic differentiation of TAMs in 2D culture.
Fig. 5: A sequential Stacks assay was performed to achieve organotypic differentiation of TAMs in 3D culture.
Fig. 6: Selected transcriptional profile of od-TAMs generated from culture with primary prostate cancer punch-biopsy samples.

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Data availability

The authors declare that all data supporting the findings of this study are available within the paper and its Supplementary Information. Source data for the primary-sample figures are available in Figshare with the identifier https://figshare.com/s/c237024425f6e06e8fcc.

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Acknowledgements

This work is funded by a National Science Foundation grant (EFRI-MKIS), University of Wisconsin Carbone Cancer Center Cancer Center Support Grant (P30 CA014520), Research Training in Hematology T32 (NIH T32 HL07899), NIH R01EB010039 BRG, NIH R01 CA185251, NIH K12 DK100022, DOD Prostate Cancer Research Program (W81XWH-16-0543), the Arnold and Mabel Beckman Foundation (Beckman Young Investigator Award), and the University of Washington. We thank D. Kosoff for helpful discussions and assistance with PCR, and undergraduate student B. Horman for facilitating experiments.

Author information

Author notes

  1. These authors contributed equally: Jiaquan Yu, Erwin Berthier.

Authors and Affiliations

  1. Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA

    Jiaquan Yu, Erwin Berthier, Alexandria Craig, Theodorus E. de Groot, Sidney Sparks, Patrick N. Ingram, David J. Beebe & Ashleigh B. Theberge

  2. Department of Chemistry, University of Washington, Seattle, WA, USA

    Erwin Berthier & Ashleigh B. Theberge

  3. Carbone Cancer Center, University of Wisconsin-Madison, Madison, WI, USA

    David F. Jarrard & David J. Beebe

  4. Department of Urology, University of Wisconsin Carbone Cancer Center, Madison, WI, USA

    David F. Jarrard

  5. Department of Pathology and Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA

    Wei Huang

  6. Department of Urology, University of Washington School of Medicine, Seattle, WA, USA

    Ashleigh B. Theberge

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Contributions

J.Y., E.B., S.S., D.J.B. and A.B.T. designed the research. J.Y., A.C. and S.S. conducted experiments; all of the authors interpreted the data. J.Y., A.B.T., E.B. and D.J.B. wrote the manuscript, and all authors revised it.

Corresponding authors

Correspondence to David J. Beebe or Ashleigh B. Theberge.

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Competing interests

The authors have potential conflicts of interest related to technologies presented here: J.Y. (Stacks to the Future), E.B. (Tasso, Salus Discovery and Stacks to the Future), T.E.d.G. (Stacks to the Future and Lynx Biosciences), A.B.T. (Stacks to the Future), and D.J.B. (Bellbrook Labs, Tasso, Stacks to the Future, Lynx Biosciences, Onexio Biosystems and Salus Discovery). However, none of these companies supported this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–6, methods and captions for Supplementary Videos 1–3.

Reporting summary

Supplementary Video 1

Stacks layers are open microfluidic devices that can be operated with a pipette.

Supplementary Video 2

Stacks devices can be assembled to enable diffusion.

Supplementary Video 3

Stacks devices are reconfigurable and enable diffusion across layers.

ImageJ macros

ImageJ macros for image processing.

Injection-moulding CAD designs

SolidWorks files for the Stack devices.

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Yu, J., Berthier, E., Craig, A. et al. Reconfigurable open microfluidics for studying the spatiotemporal dynamics of paracrine signalling. Nat Biomed Eng 3, 830–841 (2019). https://doi.org/10.1038/s41551-019-0421-4

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