Robotic fluidic coupling and interrogation of multiple vascularized organ chips

Abstract

Organ chips can recapitulate organ-level (patho)physiology, yet pharmacokinetic and pharmacodynamic analyses require multi-organ systems linked by vascular perfusion. Here, we describe an ‘interrogator’ that employs liquid-handling robotics, custom software and an integrated mobile microscope for the automated culture, perfusion, medium addition, fluidic linking, sample collection and in situ microscopy imaging of up to ten organ chips inside a standard tissue-culture incubator. The robotic interrogator maintained the viability and organ-specific functions of eight vascularized, two-channel organ chips (intestine, liver, kidney, heart, lung, skin, blood–brain barrier and brain) for 3 weeks in culture when intermittently fluidically coupled via a common blood substitute through their reservoirs of medium and endothelium-lined vascular channels. We used the robotic interrogator and a physiological multicompartmental reduced-order model of the experimental system to quantitatively predict the distribution of an inulin tracer perfused through the multi-organ human-body-on-chips. The automated culture system enables the imaging of cells in the organ chips and the repeated sampling of both the vascular and interstitial compartments without compromising fluidic coupling.

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Fig. 1: Overview of the Interrogator device.
Fig. 2: Schematic of stage calibration and system characterization.
Fig. 3: Linking scheme of eight organ HuBoCs.
Fig. 4: Automated HuBoC linkage demonstrates maintenance of organ viability and function for 3 weeks.
Fig. 5: Long-term analysis of inulin-FITC PK in an eight organ system linked via vasculature and supported by computational PBPK modelling.

Data availability

All the data supporting the results in this study are available within the article and its Supplementary Information. Mass spectrometry data of brain-chip metabolites are available as Supplementary Dataset 1. The broad range of raw datasets acquired and analysed (or any subsets of it), which for reuse would require contextual metadata, are available from the corresponding author upon reasonable request.

Code availability

A SolidWorks CAD package of the Interrogator that defines all hardware components and assembly instructions are provided as Supplementary Design Files 1 and 2. The mass spectrometry data are provided as Supplementary Dataset 1. Control software is available at https://gitlab.com/wyss-microengineering/hydra-controller, and video tutorials are available at https://vimeo.com/album/5703210. Organ-chip simulations developed using CoBi Tools are available at http://medicalavatars.cfdrc.com/index.php/cobi-tools.

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Acknowledgements

This research was sponsored by the Wyss Institute for Biologically Inspired Engineering at Harvard University and the Defense Advanced Research Projects Agency under Cooperative Agreement number W911NF-12-2-0036. The views and conclusions contained in this document are those of the authors and should not be interpreted as representing the official policies, either expressed or implied, of the Defense Advanced Research Projects Agency or the US government. This work was performed in part at the Center for Nanoscale Systems (CNS), a member of the National Nanotechnology Coordinated Infrastructure Network (NNCI), which is supported by the National Science Foundation under NSF award number 1541959. The CNS is part of Harvard University, the Harvard Materials Research Science and Engineering Center (DMR-1420570). The authors thank J. Caramanica and P. Machado for their machining expertise, M. Rosnach for his artwork, B. Fountaine and S. Kroll for their help with photography, M. Rousseau for help with videography, C. Vidoudez for mass spectrometry analysis, and J. Wikswo for helpful input at the start of this project.

Author information

R.N., A.H., B.M.M. and R.P.-B. led the data analyses for the generation of figures and, with D.E.I., prepared the manuscript. A.H., B.M.M., A. Chalkiadaki, D.B.C, M.C., T.H., M.B., S.D., E.A.F., S.S.F.J., T.G., S.J.-F., V.K., L.L., R.M., Y.M., J.A.N., B.O., T.-E.P., H.S., B.S., G.J.T., Z.T., T.H.-I., K.-J.J., A.S.-P. and M.Y. planned and performed biological experiments, with G.A.H., O.L., A.B., R.N., R.P.-B., K.K.P. and D.E.I. supervising the work. A.Cho., E.C., Y.C., J.F., R.F., C.F.N., R.N., G.A.H., J.A.G., N.W., K.K.P. and D.E.I. were responsible for the development and fabrication of the chip. J.S., G.T.II, C.H., J.F.-A., J.A.G. and D.L. conceptualized the robotics-based organ chip linking and developed early versions of the Interrogator instrument and software with an integrated robotic sampler. M.I., Y.C., S.M., A.D., T.D., T.F., O.H. B.A.N., and R.N were responsible for the software and hardware engineering and were involved in the development of the final instrument and sensors used in this study. D.D., M.R.S. and A.P. were responsible for MCRO model development and data analyses, working closely with A.H., B.M.M., R.P.-B. and R.N. Finally, R.N., R.P.-B., O.L., G.A.H., D.L., K.K.P. and D.E.I. were responsible for overseeing and orchestrating the entire effort.

Correspondence to Donald E. Ingber.

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Competing interests

D.E.I. is a founder and holds equity in Emulate, Inc., and chairs its scientific advisory board. K.K.P. is a consultant and a member of the scientific advisory board of Emulate, Inc. S.S.F.J., J.F.-A., G.A.H., C.H., K.-J.J., V.K., L.L., D.L., J.N., J.S., G.T.II and N.W. are employees of and hold equity in Emulate, Inc. A.B., Y.C., M.C., S.D., J.F.-A., T.F., E.A.F., J.A.G., G.A.H., T.H.-I., O.H., A.H., C.H., D.E.I., M.I., K.-J.J., V.K., L.L., D.L., O.L., B.M.M., Y.M., J.N., R.N., T.-E.P., K.K.P., J.S., A.S.-P., G.T.II and N.W. are inventors on intellectual property licensed to Emulate, Inc.

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Supplementary information

41551_2019_497_MOESM6_ESM.mp4

Overview of the Interrogator-device components and organ-chip linking.

41551_2019_497_MOESM7_ESM.mov

Gut-chip stretching, visualized via the microscope module.

41551_2019_497_MOESM8_ESM.mov

Heart chip beating after being linked for 3 weeks on the Interrogator device.

41551_2019_497_MOESM9_ESM.avi

High‐fidelity diffusion model of the heart chip.

Supplementary Information

Supplementary methods, figures, tables and video captions.

Reporting Summary

Supplementary Design Files 1

SolidWorks CAD package for the interrogator device.

Supplementary Design Files 2

SolidWorks CAD package for the fluorescence module.

Supplementary Dataset 1

Mass spectrometry data of brain-chip metabolites.

Supplementary Video 1

Overview of the Interrogator-device components and organ-chip linking.

Supplementary Video 2

Gut-chip stretching, visualized via the microscope module.

Supplementary Video 3

Heart chip beating after being linked for 3 weeks on the Interrogator device.

Supplementary Video 4

High‐fidelity diffusion model of the heart chip.

Supplementary Video 5

CoBi Q‐3D model of gut–liver chip linking.

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Novak, R., Ingram, M., Marquez, S. et al. Robotic fluidic coupling and interrogation of multiple vascularized organ chips. Nat Biomed Eng (2020). https://doi.org/10.1038/s41551-019-0497-x

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