SpringerLink
Log in

Microfluidic 3D Cytotoxic Assay

  • Protocol
  • First Online:
Microfluidics Diagnostics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2804))

  • 220 Accesses

Abstract

Microfluidic-based cytotoxic assays provide high physiological relevance with the potential to replace conventional animal experiments and two-dimensional (2D) assays. Here, a 3D method utilizing a microfluidic platform for analysis of lymphocyte cytotoxicity is introduced in detail, including platform design, cell culture method, real-time cytotoxic assay setup, and image-based analysis. A 2D experimental method is used for comparison, which effectively demonstrates the advantages of 3D microfluidic platforms in closely recapitulating immune responses within the tumor microenvironment. Moreover, a wide range of experimental possibilities and applications using microfluidic 3D cytotoxic assays is introduced in this chapter, along with their capabilities, limitations, and future outlook.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
EUR 44.95
Price includes VAT (Spain)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
EUR 169.99
Price includes VAT (Spain)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
EUR 207.99
Price includes VAT (Spain)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Galon J, Bruni D (2020) Tumor immunology and tumor evolution: intertwined histories. Immunity 52:55–81

    Article  CAS  PubMed  Google Scholar 

  2. Zhang J et al (2022) Immunotherapy discovery on tumor organoid-on-a-chip platforms that recapitulate the tumor microenvironment. Adv Drug Deliv Rev 187:114365

    Article  CAS  PubMed  Google Scholar 

  3. Bader JE, Voss K, Rathmell JC (2020) Targeting metabolism to improve the tumor microenvironment for cancer immunotherapy. Mol Cell 78:1019–1033

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Buoncervello M, Gabriele L, Toschi E (2019) The Janus face of tumor microenvironment targeted by immunotherapy. Int Journal of Mol Sci 20:4320

    Article  CAS  Google Scholar 

  5. Khalil DN et al (2016) The future of cancer treatment: immunomodulation, CARs and combination immunotherapy. Nat Rev Clin Oncol 13:273–290

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Sterner RC, Sterner RM (2021) CAR-T cell therapy: current limitations and potential strategies. Blood Cancer J 11:69

    Article  PubMed  PubMed Central  Google Scholar 

  7. Giraldo NA et al (2019) The clinical role of the TME in solid cancer. Br J Cancer 120:45–53

    Article  PubMed  Google Scholar 

  8. Rongvaux A et al (2014) Development and function of human innate immune cells in a humanized mouse model. Nat Biotechnol 32:364–372

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Vesely MD et al (2011) Natural innate and adaptive immunity to cancer. Annu Rev Immunol 29:235–271

    Article  CAS  PubMed  Google Scholar 

  10. Tseng D et al (2013) Anti-CD47 antibody mediated phagocytosis of cancer by macrophages primes an effective antitumor T-cell response. Proc of Nat Acad of Sci 110:11103–11108

    Article  CAS  Google Scholar 

  11. Hirt C et al (2014) In vitro 3D models of tumor-immune system interaction. Adv Drug Deliv Rev 79-80:145–154

    Article  CAS  PubMed  Google Scholar 

  12. Sontheimer-Phelps A, Hassell BA, Ingber DE (2019) Modelling cancer in microfluidic human organs-on-chips. Nat Rev Cancer 19(2):65–81

    Article  CAS  PubMed  Google Scholar 

  13. Park D et al (2019) High-throughput microfluidic 3D cytotoxicity assay for cancer immunotherapy (CACI-IMPACT platform). Front Immunol 10:1133

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Park D et al (2021) Aspiration-mediated hydrogel micropatterning using rail-based open microfluidic devices for high-throughput 3D cell culture. Sci Rep 11:19986

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Ronteix G et al (2022) High resolution microfluidic assay and probabilistic modeling reveal cooperation between T cells in tumor killing. Nat Commun 13:3111

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Song J et al (2021) High-throughput 3D in vitro tumor vasculature model for real-time monitoring of immune cell infiltration and cytotoxicity. Front Immunol 12:733317

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Pavesi A et al (2015) Using microfluidics to investigate tumor cell extravasation and T-cell immunotherapies. Annu Int Conf IEEE Eng Med Biol Soc 2015:1853–1856

    CAS  PubMed  Google Scholar 

  18. Nguyen M et al (2018) Dissecting effects of anti-cancer drugs and cancer-associated fibroblasts by on-chip reconstitution of immunocompetent tumor microenvironments. Cell Rep 25:3884–3893.e3

    Article  CAS  PubMed  Google Scholar 

  19. Businaro L et al (2013) Cross talk between cancer and immune cells: exploring complex dynamics in a microfluidic environment. Lab Chip 13:229–239

    Article  CAS  PubMed  Google Scholar 

  20. Beckwith AL, Velásquez-García LF, Borenstein JT (2019) Microfluidic model for evaluation of immune checkpoint inhibitors in human tumors. Adv Healthc Mater 8:e1900289

    Article  PubMed  Google Scholar 

  21. Cui X et al (2020) Dissecting the immunosuppressive tumor microenvironments in glioblastoma- on-a-chip for optimized PD-1 immunotherapy. elife 9:e52253

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Fang T et al (2019) Remodeling of the tumor microenvironment by a chemokine/anti-PD-L1 nanobody fusion protein. Mol Pharm 16:2838–2844

    Article  CAS  PubMed  Google Scholar 

  23. Biselli E et al (2017) Organs on chip approach: a tool to evaluate cancer-immune cells interactions. Sci Rep 7:1–12

    Article  CAS  Google Scholar 

  24. Aung A et al (2020) An engineered tumor-on-a-chip device with breast cancer-immune cell interactions for assessing T-cell recruitment. Cancer Res 80:263–275

    Article  CAS  PubMed  Google Scholar 

  25. Lee SWL et al (2018) Characterizing the role of monocytes in T cell cancer immunotherapy using a 3D microfluidic model. Front Immunol 9:416

    Article  PubMed  PubMed Central  Google Scholar 

  26. Ayuso JM et al (2021) Microfluidic tumor-on-a-chip model to evaluate the role of tumor environmental stress on NK cell exhaustion, science. Advances 7:eabc2331

    CAS  Google Scholar 

  27. Ke L-Y et al (2018) Cancer immunotherapy μ-environment LabChip: taking advantage of optoelectronic tweezers. Lab Chip 18:106–114

    Article  CAS  Google Scholar 

  28. Surendran V et al (2021) A novel tumor-immune microenvironment (TIME)-on-chip mimics three dimensional neutrophil-tumor dynamics and neutrophil extracellular traps (NETs)-mediated collective tumor invasion. Biofabrication 13:035029

    Article  CAS  Google Scholar 

  29. Bi Y et al (2020) Tumor-on-a-chip platform to interrogate the role of macrophages in tumor progression. Integr Biol (Camb) 12:221–232

    Article  PubMed  Google Scholar 

  30. Ayuso JM et al (2019) Evaluating natural killer cell cytotoxicity against solid tumors using a microfluidic model. Onco Targets Ther 8:1553477

    Google Scholar 

  31. Parlato S et al (2017) 3D microfluidic model for evaluating immunotherapy efficacy by tracking dendritic cell behaviour toward tumor cells. Sci Rep 7:1–16

    Article  Google Scholar 

  32. Parlato S et al (2021) Tumor-on-a-chip platforms to study cancer–immune system crosstalk in the era of immunotherapy. Lab Chip 21:234–253

    Article  CAS  PubMed  Google Scholar 

  33. Kim S et al (2021) Microfluidic tumor vasculature model to recapitulate an endothelial immune barrier expressing FasL. ACS Biomater Sci Eng 7:1230–1241

    Article  CAS  PubMed  Google Scholar 

  34. Miccoli B, Braeken D, Li Y-CE (2018) Brain-on-a-chip devices for drug screening and disease modeling applications. Curr Pharm Des 24:5419–5436

    Article  CAS  PubMed  Google Scholar 

  35. Iyer V et al (2022) Advancing microfluidic diagnostic chips for clinical use. Lab Chip 22:3110–3121

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Ren K, Zhou J, Wu H (2013) Materials for microfluidic chip fabrication. Acc Chem Res 46:2396–2406

    Article  CAS  PubMed  Google Scholar 

  37. Van Meer B et al (2017) Small molecule absorption by PDMS in the context of drug response bioassays. Biochem Biophys Research Comm 482:323–328

    Article  Google Scholar 

  38. Leung CM et al (2022) A guide to the organ-on-a-chip. Nat Rev Methods Primers 2:33

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. The authors Hyeri Choi, Sunghun Cheong, and Ailian ** have contributed equally to this chapter

Authors and Affiliations

  1. Interdisciplinary Program in Bioengineering, Seoul National University, Seoul, South Korea

    Hyeri Choi, Sunghun Cheong, Ailian ** & Noo Li Jeon

  2. Department of Mechanical Engineering, Seoul National University, Seoul, South Korea

    Dohyun Park & Noo Li Jeon

  3. Institute of Advanced Machines and Design (SNU-IAMD), Seoul National University, Seoul, South Korea

    Noo Li Jeon

Authors
  1. Hyeri Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sunghun Cheong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ailian **
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dohyun Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Noo Li Jeon
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Noo Li Jeon .

Editor information

Editors and Affiliations

  1. Centre de recherche des Cordeliers, INSERM, UMR-S1138, CNRS SNC 5096, Université Paris Cité, Paris, Paris, France

    Valérie Taly

  2. UMR 168, Institut Pierre-Gilles de Gennes, Institut Curie, Paris, Paris, France

    Stéphanie Descroix

  3. CEA, INRAE, Médicaments et Technologies pour la Santé (MTS), SPI, Université Paris-Saclay, Gif-sur-Yvette, Paris, France

    Karla Perez-Toralla

Rights and permissions

Reprints and permissions

Copyright information

© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Choi, H., Cheong, S., **, A., Park, D., Jeon, N.L. (2024). Microfluidic 3D Cytotoxic Assay. In: Taly, V., Descroix, S., Perez-Toralla, K. (eds) Microfluidics Diagnostics. Methods in Molecular Biology, vol 2804. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3850-7_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3850-7_13

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3849-1

  • Online ISBN: 978-1-0716-3850-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation