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
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
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)
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.
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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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