Three-dimensional (3D) heterotypic microtissues

3D heterotypic microtissues composed of tumor cells, fibroblasts and immune cells provide additional information on the effects of immunotherapies compared to classical 2D systems.

3D models are a versatile platform, suitable for testing engineered immunoagents like antibodies with enhanced effector function, immunocytokines, T-cell binders and immune checkpoint modulators in monotherapy and in combination as well as cytotoxic compounds. They enable long-term co-culture with immune cells and characterization of immune cell subpopulations opening up new ways to assess the enhancement of NK cell, T-cell and NKT cell recruitment and activity (cytokine/chemokine release, infiltration, surface and intracellular marker expression, elimination of target cells and proliferation).

Advantages over 2D systems

Antibody penetration, immune cell infiltration, antibody targeting and subsequent lymphocyte recruitment and elimination of tumor- or fibroblast- microtissue areas represent additional aspects of the activity of cancer immunotherapy agents that cannot be assessed in a 2D system. Additionally, effects of combination therapies can be better evaluated in 3D systems, since such models better support a physiological immunomodulatory function than 2D systems.

Advantages over in-vivo models

There is a considerable temporal advantage over the in-vivo models because 3D models allow a faster kinetic analysis of therapeutic activity due to fast and efficient immune cell activation and target cell lysis.

Our services

IBR Inc. scientists have vast experience in 3D heterotypic microtissues models for testing cytotoxic compounds and immunotherapy agents. Our services for 3D cell-based assays include:

  • Custom assay development of 3D co-culture models
  • Hanging–drop as well as ultra-low attachment microwell platforms
  • 3D immunoassays to assess immune cell recruitment and activation
  • 3D cell-based assays to assess cytotoxicity
  • 3D cell-based assays to assess antibody penetration and targeting
  • Extensive choice of assay readouts
    We are experienced in assessing the results of 3D assays though a variety of readouts for a comprehensive assessment of the mechanisms of action of your molecule in monotherapy and in combination. Our readouts include:
    • IHC and quantitative image analysis
    • Analysis of supernatants (cytometric bead array for quantitation of cytokines, LDH measurement for quantitation of target cell death)
    • ATP measurement for quantification of cell proliferation
    • Confocal microscopy and immunofluorescence for assessment of antibody penetration and targeting
    • Live microscopy for monitoring microtissue compaction and growth
    • Microscopic assessment of microtissue size
    • Flow cytometry analysis, upon microtissue dissociation, for cell phenotyping
  • Quantitative image analysis
    IBR Inc. scientists are able to extract high-level information and have vast experience in quantitative image analysis for the evaluation of immunohistochemical staining of 3D cell cultures. Quantification of immune cell infiltration, tumor and stromal areas is of essential importance for the evaluation of complex 3D immunoassays, in order to assess the effects of immunotherapic agents.

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