In vitro assay development services

2D and 3D cell based assay development in oncology and immuno-oncology

At Oncodesign, we offer specifically adapted cellular assays to suit project’s needs. Assays can be set-up to characterize complex phenotypes. Multiple end-points can be assessed within the same sample. More complex, lower throughput, more physiological plate-based assays can be conducted, generally in cells relevant to the target mechanism of disease.

Representation of Immunogenic cell death inducer | Oncodesign

Immunogenic cell death (ICD) inducer

Cancer cells subjected to some type of chemotherapeutics and other anti-cancer targeted therapies:

  • Expose calreticulin and other endoplasmic reticulum chaperones on their surface,
  • Secrete ATP (adenosine triphosphate),
  • Initiate type I interferon (IFN) response,
  • Release high-mobility group box 1 (HMGB1) and annexin A1 (ANXA1).

When secreted or exposed extracellularly, they bind to their cognate receptors on the surface of myeloid or lymphoid cells, which enables the engulfment of cell corpses by antigen-presenting cells, including dendritic cells (DCs). This process in the context of proper immunostimulatory signals, eventually leads to the priming of an adaptive immune response. This culminates in the establishment of a cytotoxic T lymphocytes (CTL)-mediated anti-cancer immune response with potential to kill therapy-resistant cancer cells via an IFN-γ-dependent mechanism.

Depending on the cancer type, this could be correlated with good or bad prognosis, as well as markers of an active anti-cancer immune response.​

In combination with our medicinal chemists, in vivo pharmacologists and DMPK scientists, our integrated team can accelerate your research program by an understanding and determination of the mechanism of action. Learn more about our integrated services:

  • INPACT: integrated service in preclinical pharmacology
  • DRIVE-IDDS: integrated drug discovery solutions to generate novel drug candidates

Let’s talk about your project to know how we can best assist you.


DC activation (co-culture) assay

In this example, DiO-labeled CT26 cells were treated with the indicated concentrations of dinaciclib (DIN), a CDK inhibitor, for 24 hours and then co-cultured with mDCs for an additional 24 hours. The percentage of CD11c+ DCs with engulfed tumor cells was assessed by flow cytometry. Secretion of IL-1β and IL-6 into the co-culture supernatant was determined by multiplex assay.​


Dinaciclib-treated tumor cells enhance DC function | Oncodesign

Legend: Dinaciclib-treated tumor cells enhance DC function. Left: phagocytosis of DIN-treated CT26 cells by DCs; right: secretion of cytokines in co-culture supernatant.

Dinaciclib-treated tumor cells were efficiently phagocytosed by DCs (left graph), resulting in increased DC maturation, as indicated by the surface expression of MHCII, CD80, and CD86 (not shown). We also found increased secretion of IL-1β (right graph) in the co-culture supernatant. Secretion of IL-1β from DCs in response to purinergic receptor agonists (ATP) and TLR4 ligands (HMGB1) plays an important role in antitumor T cell priming. In addition to DC activation and T cell priming assays, Oncodesign has the capacity to perform medium-throughput screen to identify damage-associated molecular pattern (DAMPs) in the context of ICD.

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3D T cell cytotoxicity assay

Physiologically relevant assays to monitor the complex biology in in vitro assays are the need of the day. Killing of unwanted target cells, such as tumor cell, is a critical component of the human host defense mechanism. T cell-mediated killing is one of the cell-mediated immune response mechanism. It involves the stimulation of CTL that then actively lyse target cells. T cell cytotoxicity assay is an in vitro assay that can be used to assess immune enhancement or immune suppression by therapeutic antibodies, T cell therapies and small molecules. Our team can customize the routine immuno-oncology T cell assays to suit your needs.

In this example, HCT116 cells were stably transfected with Incucyte® Nuclight Red Lentivirus reagent (EF-1 Alpha promoter, Puromycin selection) to enable live cell imaging and monitoring over time. The cells were grown in ultra-low attachment plates to enable spheroid formation.

3D Spheroid T cell cytotoxicity assay | Oncodesign
T cell cytoxicity assay visualized by imaging

Legend: PBMC-mediated tumor killing in 3D format.

Upon addition of PKH67 labelled human peripheral blood mononuclear cells (PBMCs) in the absence of any stimulation, the PBMCs did not induce tumor cell killing (gray bar). Upon stimulation of the PBMCs with anti-CD3 and IL-2, a reduction in red fluorescence due to the induction of tumor cell killing was observed (green bar).​

To support your research efforts in oncology and immuno-oncology, our integrated and customized package INPACT PoC Oncology contains all the requirement for a successful preclinical program.

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