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OcellO offers compound testing services using co-cultures of cancer and immune cells.  These test the ability of compounds to potentiate infiltration of T cells into the tumor and enhance their cytotoxic activity.  Diverse cultured tumor tissues, including tumoroids derived from established tumor cell lines, colorectal cancer organoids and PDX material, can be used in these assays. Tumoroids are co-cultured with partially HLA-matched PBMCs from healthy donors, purified T cell populations (e.g. CD8+, CD3+), engineered T cells, CAR T cells or myeloid cells differentiated in vitro (e.g. DCs, M1 and M2 macrophages).

A suite of assays with a flexible design

Assays can be customized by replacing any of the cellular players. This allows for testing of a diverse range of immunotherapies that focus on different immune compartments and target diverse cancer indications.

Representative images of subsequent steps occurring during tumoroid-T cells encounter

Effects of drugs that aim at enhancing any of these steps can be visualized and quantified in a robust and high throughput manner.

Example Data

Functional endpoints: T cell infiltration and tumor killing

Functional endpoints: T cell infiltration and tumor killing

Breast cancer cells were seeded in 3D to form tumoroids. CD3+ T cells isolated  from PBMCs were added with immunomodulators. Immune cells and cancer cells were stained separately and imaged. Automated image analysis measured infiltration of immune cells into  tumoroids and tumor cell killing Treatment effects were quantified.

Determination of intermediate stages of immune cells activation

Determination of intermediate stages of immune cells activation

Differentially pre-treated PBMCs co-cultured with SKBR- 3 tumoroids. Bi-parametric analysis of tumoroids size vs T cells invasiveness enables a better understanding of a drugs immunomodulatory profile: do they impact on infiltration/killing or both?

Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC)

Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC)

Her2– and Her2+ breast cancer cell lines were seeded in 3D to form tumoroids. PBMCs were added and cultured further in the presence of Trastuzumab. Cultures were fixed, stained and imaged. Image-based analysis was applied to measure immune cell-mediated killing. Enhanced killing induced by Trastuzumab was quantified.

Key advantages

  • Automated analysis and robust quantification of activity of immune cells
  • Functional read-outs:  active migration, infiltration into tumoroids, tumor cell killing
  • Physiologically relevant 3D micro-environment
  • Visualization of immune cell  interaction with the tumor
  • HLA-matched cell types

Assay Principle

3D co-cultures are optimized for study requirements. Tumor cells are grown embedded in a 3D extracellular matrix protein-rich hydrogel; immune cells are added together with test compounds and co-cultures are maintained for 1-4 days. Immune  cells are stained separately to allow for distinction from cancer cells.  After ‘optical sectioning’ 3D image stacks are reconstituted. Robust  high-throughput (384 well) screening is done using image-based  measurements of selected features: T cell invasiveness, total tumor  volume, shape and size of tumoroids.

Co-culture

A unique screening platform for efficient testing of treatment effects in assays that yield functional data on the tumor – immune systeminterplay.

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    Find us here

    OcellO B.V.
    J.H. Oortweg 21
    2333 CH Leiden
    The Netherlands

    Email [email protected]
    Telephone +31 (0)71 3322 875 
    Chamber of Commerce
    (KvK) number: 53055926