| Autor: |
De Ninno A; CNR Institute for Photonics and Nanotechnology; adele.deninno@cnr.it., Bertani FR; CNR Institute for Photonics and Nanotechnology., Gerardino A; CNR Institute for Photonics and Nanotechnology., Schiavoni G; Dept. of Oncology and Molecular Medicine, Istituto Superiore di Sanità., Musella M; Istituto di Patologia Generale, Università Cattolica del Sacro Cuore., Galassi C; Istituto di Patologia Generale, Università Cattolica del Sacro Cuore., Mattei F; Dept. of Oncology and Molecular Medicine, Istituto Superiore di Sanità., Sistigu A; Istituto di Patologia Generale, Università Cattolica del Sacro Cuore; Tumor Immunology and Immunotherapy Unit, IRCCS Regina Elena National Cancer Institute; antonella.sistigu@gmail.com., Businaro L; CNR Institute for Photonics and Nanotechnology. |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of visualized experiments : JoVE [J Vis Exp] 2021 Apr 30 (170). Date of Electronic Publication: 2021 Apr 30. |
| DOI: |
10.3791/61895 |
| Abstrakt: |
Complex disease models demand cutting-edge tools able to deliver physiologically and pathologically relevant, actionable insights, and unveil otherwise invisible processes. Advanced cell assays closely mimicking in vivo scenery are establishing themselves as novel ways to visualize and measure the bidirectional tumor-host interplay influencing the progression of cancer. Here we describe two versatile protocols to recreate highly controllable 2D and 3D co-cultures in microdevices, mimicking the complexity of the tumor microenvironment (TME), under natural and therapy-induced immunosurveillance. In section 1, an experimental setting is provided to monitor crosstalk between adherent tumor cells and floating immune populations, by bright field time-lapse microscopy. As an applicative scenario, we analyze the effects of anti-cancer treatments, such as the so-called immunogenic cancer cell death inducers on the recruitment and activation of immune cells. In section 2, 3D tumor-immune microenvironments are assembled in a competitive layout. Differential immune infiltration is monitored by fluorescence snapshots up to 72 h, to evaluate combination therapeutic strategies. In both settings, image processing steps are illustrated to extract a plethora of immune cell parameters (e.g., immune cell migration and interaction, response to therapeutic agents). These simple and powerful methods can be further tailored to simulate the complexity of the TME encompassing the heterogeneity and plasticity of cancer, stromal and immune cells subtypes, as well as their reciprocal interactions as drivers of cancer evolution. The compliance of these rapidly evolving technologies with live-cell high-content imaging can lead to the generation of large informative datasets, bringing forth new challenges. Indeed, the triangle ''co-cultures/microscopy/advanced data analysis" sets the path towards a precise problem parametrization that may assist tailor-made therapeutic protocols. We expect that future integration of cancer-immune on-a-chip with artificial intelligence for high-throughput processing will synergize a large step forward in leveraging the capabilities as predictive and preclinical tools for precision and personalized oncology. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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