Tumor Organoid and CAR-NK Co-Culture Model for Immuno-Oncology Drug Discovery and Development

NK cells pose a very low risk of causing graft-versus-host disease when used in allogeneic therapies, making CAR-NK cells a promising off-the-shelf product that can be used in allogeneic therapies. Creative Biolabs has capitalized on this property of CAR-NK cells and combined it with a 3D ex vivo bio-platform to provide a co-culture model to increase therapeutic accessibility.

Preparation of Expanded Multi-Source CAR-NK Cells

In addition to autologous-derived NK cells, other sources that can be used for CAR-NK include peripheral blood, umbilical cord blood, stem cell differentiation, and NK cell lines.

Peripheral blood source.

CAR-NK cells prepared from peripheral-derived NK cells will usually be dominated by CD56dimCD16+ NK cells, which can account for more than 90% of the total number of NK cells, and this class of NK cells has a strong toxic effect but its proliferation ability is weak.

Cord blood source.

Like peripheral blood-derived NK cells, NK cells from umbilical cord blood can be enriched with high purity by sorting kits. Cord blood-derived NK cells tend to exhibit an immature phenotype and low cytotoxicity.

Stem cell HSC/iPSC-induced source.

It is expected to address the defect that peripheral blood and cord blood-derived NK are difficult to standardize. HSC/iPSC has infinite passaging properties and genetic editability, enabling the preparation of CAR-HSC/iPSC. As a stable source, CAR-HSC/iPSC continuously prepares highly homogeneous CAR-NK cells with predominantly immature NK cells.

NK cell line source.

CAR expression of NK cells was achieved by lentiviral or retroviral transfection, and radiation was administered to limit their in vivo lifespan before administration.

Fig. 1 CAR-NK sources and characteristics.^1,3

Co-culture Modeling and Testing

• Targeted selection of organoids of interest or specific mutations based on gene or protein expression characteristics.
• Pre-labeled CAR-NK cells are added to the co-culture system.
• Model volumes are collected at different time points to assess the effect of the intervention and differential immune response over time. Available assays include but are not limited to, morphological evaluation, reporter gene analysis, live-dead cell flow assays, and cytokine analysis.

Application Advantages

• Organoids are more clinically relevant compared to cancer cell lines and simpler compared to humanized in vivo systems.
• Spatial construction of multiple cell lineages to form co-culture organoids for different patients and cancer types, better reflecting the diversity and response predictive features in immune drug screening.
• Use co-culture models to combine test drugs and evaluate any potential off-target effects prior to using more complex in vivo models.
• High-inclusion techniques are applied to such co-culture studies to accurately determine the effects of drugs on physiologically relevant morphological features,  such as tumor cell killing, growth arrest, invasion, and immune cell proliferation.

Fig 2. Schematic diagram of CAR-NK exploration experiments in different solid tumors.^2,3

Taking advantage of the less toxic and less severe adverse effects of CAR-NK cell therapy and the high degree of simulation of 3D tumor models, Creative Biolabs provides co-culture models of both to help revolutionize our customers' immune drug development. Please contact us to get a solution.

Related Sections:

• Tumor Organoid Monoculture Model for Immuno-Oncology Drug Discovery and Development
• Tumor Organoid and Immune Cell Co-culture Model for Immuno-Oncology Drug Discovery and Development