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Multiscale cytometry and regulation of 3D cell cultures on a chip

Author

Listed:
  • Sébastien Sart

    (Ecole Polytechnique, CNRS-UMR7646)

  • Raphaël F.-X. Tomasi

    (Ecole Polytechnique, CNRS-UMR7646)

  • Gabriel Amselem

    (Ecole Polytechnique, CNRS-UMR7646)

  • Charles N. Baroud

    (Ecole Polytechnique, CNRS-UMR7646)

Abstract

Three-dimensional cell culture is emerging as a more relevant alternative to the traditional two-dimensional format. Yet the ability to perform cytometry at the single cell level on intact three-dimensional spheroids or together with temporal regulation of the cell microenvironment remains limited. Here we describe a microfluidic platform to perform high-density three-dimensional culture, controlled stimulation, and observation in a single chip. The method extends the capabilities of droplet microfluidics for performing long-term culture of adherent cells. Using arrays of 500 spheroids per chip, in situ immunocytochemistry and image analysis provide multiscale cytometry that we demonstrate at the population scale, on 104 single spheroids, and over 105 single cells, correlating functionality with cellular location within the spheroids. Also, an individual spheroid can be extracted for further analysis or culturing. This will enable a shift towards quantitative studies on three-dimensional cultures, under dynamic conditions, with implications for stem cells, organs-on-chips, or cancer research.

Suggested Citation

  • Sébastien Sart & Raphaël F.-X. Tomasi & Gabriel Amselem & Charles N. Baroud, 2017. "Multiscale cytometry and regulation of 3D cell cultures on a chip," Nature Communications, Nature, vol. 8(1), pages 1-13, December.
  • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_s41467-017-00475-x
    DOI: 10.1038/s41467-017-00475-x
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    Cited by:

    1. Yangteng Ou & Shixiang Cao & Yang Zhang & Hongjia Zhu & Chengzhi Guo & Wei Yan & Fengxue Xin & Weiliang Dong & Yanli Zhang & Masashi Narita & Ziyi Yu & Tuomas P. J. Knowles, 2023. "Bioprinting microporous functional living materials from protein-based core-shell microgels," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
    2. Gustave Ronteix & Shreyansh Jain & Christelle Angely & Marine Cazaux & Roxana Khazen & Philippe Bousso & Charles N. Baroud, 2022. "High resolution microfluidic assay and probabilistic modeling reveal cooperation between T cells in tumor killing," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    3. L. Mathur & B. Szalai & N. H. Du & R. Utharala & M. Ballinger & J. J. M. Landry & M. Ryckelynck & V. Benes & J. Saez-Rodriguez & C. A. Merten, 2022. "Combi-seq for multiplexed transcriptome-based profiling of drug combinations using deterministic barcoding in single-cell droplets," Nature Communications, Nature, vol. 13(1), pages 1-15, December.

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