Article | Published:

Reconfigurable microfluidic hanging drop network for multi-tissue interaction and analysis

Nature Communications volume 5, Article number: 4250 (2014) | Download Citation

Abstract

Integration of multiple three-dimensional microtissues into microfluidic networks enables new insights in how different organs or tissues of an organism interact. Here, we present a platform that extends the hanging-drop technology, used for multi-cellular spheroid formation, to multifunctional complex microfluidic networks. Engineered as completely open, ‘hanging’ microfluidic system at the bottom of a substrate, the platform features high flexibility in microtissue arrangements and interconnections, while fabrication is simple and operation robust. Multiple spheroids of different cell types are formed in parallel on the same platform; the different tissues are then connected in physiological order for multi-tissue experiments through reconfiguration of the fluidic network. Liquid flow is precisely controlled through the hanging drops, which enable nutrient supply, substance dosage and inter-organ metabolic communication. The possibility to perform parallelized microtissue formation on the same chip that is subsequently used for complex multi-tissue experiments renders the developed platform a promising technology for ‘body-on-a-chip’-related research.

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Acknowledgements

This work was financially supported by the FP7 of the EU through the project ‘Body on a chip’, ICT-FET-296257 and an individual Ambizione Grant 142440 of the Swiss National Science Foundation for Olivier Frey. We would like to thank Thomas Horn of the D-BSSE Single Cell Unit at ETH Zurich for microscopy assistance. InSphero AG, Switzerland, is acknowledged for providing primary rat liver cell suspensions and culture media.

Author information

Affiliations

  1. ETH Zürich, Department of Biosystems Science and Engineering, Bio Engineering Laboratory, Mattenstrasse 26, CH-4058 Basel, Switzerland

    • Olivier Frey
    • , Patrick M. Misun
    •  & Andreas Hierlemann
  2. InSphero AG, Wagistrasse 27, CH-8952 Schlieren, Switzerland

    • David A. Fluri
  3. Leibniz Research Centre for Working Environment and Human Factors (IfADo), TU Dortmund University, Ardeystrasse 67, D-44139 Dortmund, Germany

    • Jan G. Hengstler

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Contributions

O.F. conceived the hanging drop network concept, designed and fabricated the microfluidic devices and performed experiments, analysed data and wrote the manuscript; P.M.M. designed and fabricated the microfluidic devices, performed experiments and analysed data; D.A.F. helped to design bio-activation experiments and provided primary cell preparations; J.G.H. proposed the concept for the bio-activation experiments; A.H. wrote and edited the manuscript and was involved in the scientific considerations. All authors discussed the results and implications and commented on the manuscript at all stages.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Olivier Frey.

Supplementary information

PDF files

  1. 1.

    Supplementary Figures, Supplementary Table and Supplementary Notes

    Supplementary Figures 1-9, Supplementary Table 1, Supplementary Notes 1-2

Videos

  1. 1.

    Supplementary Movie 1

    Time-lapse micrographs (phase contrast and green-fluorescence overlay, 12 fps, acquisition interval 15 min) of parallel spheroid formation based on HTC-116 eGFP colon cancer cells using different initial cell concentrations per column (15,000, 25,000, 40,000 and 70,000 cells ml-1). The respective diameters 60 hours after seeding are 172±15 μm, 194±11 μm, 226±8 μm and 284±13 μm. Scale bar is 200 μm.

  2. 2.

    Supplementary Movie 2

    Parallel bio-activation experiment. Complete continuous time-lapse movie (10 days, 30 min acquisition interval, 12 fps) of first, primary rat liver (rLiMT) and HCT-116 eGFP cancer spheroid formation (86 hours) and, second, closed-loop perfusion with different cyclophosphamide concentrations (CP; 0.0 mM, 0.1 mM and 1.0 mM) for 154 hours. CP is a pro-drug, which only develops its cytostatic effect on cancer, when first metabolized in the liver. rLiMT were not imaged during drug activation. Scale bar is 200 μm.

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DOI

https://doi.org/10.1038/ncomms5250

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