General Information

MDSC-targeted Platform

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Myeloid-derived suppressor cells (MDSCs) are key mediators of cancer-associated immunosuppression and therapy resistance, contributing to tumor progression by suppressing T-cell function within the tumor microenvironment. Their accumulation and trafficking into tumors are regulated by tumor- and stroma-derived signals, including those from cancer-associated fibroblasts.
We developed a Transwell-based co-culture platform that integrates MDSC-like cells with tumor organoids to quantitatively assess MDSC trafficking and infiltration. Combined with functional co-culture assays, this system enables evaluation of MDSC-mediated immunosuppression and supports testing of MDSC-targeted anticancer agents in a multicellular context.
Organism
Product Type

Tissue

Disease

Applications

  • Cancer Organoid: Cancer Organoid, Colorectal Cancer, Non-Small Cell Lung Cancer, Pancreatic Cancer, Breast Cancer, Cancer Organoid, Cholangiocarcinoma, Gastric Cancer, Ovarian Cancer

Principle

Chemotactic recruitment of MDSCs

MDSCs migrate toward tumor-derived signals and infiltrate tumor tissues through extracellular matrix barriers. The Transwell system models this process by enabling measurement of directional migration and invasion toward tumor organoids.

Immunosuppressive Function
MDSCs suppress T-cell activation and clustering through both direct cell–cell interactions and soluble factors. Functional co-culture assays enable quantification of this immunosuppressive activity.

Process

Generation and Phenotypic Validation of MDSC-like Cells
THP-1 cells are first differentiated under cytokine-driven conditions to generate MDSC-like cells. These cells are validated to exhibit an MDSC-consistent phenotype through flow cytometry and to express immune-regulatory gene signatures as confirmed by qRT–PCR, while remaining distinct from conventional macrophage differentiation states.
Functional Validation of Immunosuppressive Activity
Their immunosuppressive function is then assessed using PBMC and T-cell co-culture assays, where MDSC-like cells demonstrate the ability to suppress T-cell clustering through both contact-dependent interactions and soluble factor–mediated mechanisms.
Transwell Co-culture Setup
For the co-culture system, MDSC-like cells are seeded in the upper chamber of a Matrigel-coated Transwell insert with a 5 µm pore size, while colorectal cancer (CRC) organoids are placed in the lower chamber. This configuration enables the assessment of MDSC trafficking and infiltration under defined multicellular conditions.
Compound Treatment and Evaluation

Candidate compounds can be introduced into the system to evaluate their effects on MDSC behavior, including both direct cellular responses and changes in migration and infiltration within the co-culture setup.

Step 0
THP-1
Human cell line
Step 01
Standardized MDSC-like cells
Cytokine differentiation Validated by FACS + PCR
Step 02
Immune suppression assay​
T-cell co-culture Contact + soluble factors
Step 03
Drug screening in tumor model​
CRC organoid + CAF Quantified infiltration Imaging + FACS

Read out

MDSC immunosuppressive function is assessed through T-cell clustering assays, while trafficking and infiltration are quantitatively measured using imaging-based cell counting and flow cytometry (FACS), providing orthogonal and complementary readouts. The platform demonstrates that tumor organoids contribute to MDSC infiltration dynamics, while MDSC-like cells exhibit robust immunosuppressive activity in co-culture systems.

In drug profiling studies, differential responses are observed across tumor organoids and MDSC-like cells enabling evaluation of cell type–specific effects. Importantly, certain compounds reduce MDSC-mediated immunosuppression, while others modulate MDSC trafficking and infiltration in a concentration-dependent manner, highlighting distinct and complementary therapeutic strategies.

Implications for Drug Development

This MDSC-targeted Anticancer Drug Evaluation Platform (Infiltration) enables:

  • Mechanism-driven evaluation of MDSC-targeting therapies
  • Identification of agents that disrupt immune suppression via trafficking inhibition
  • More predictive assessment of drug efficacy in a human, multicellular tumor cont
Ultimately, it supports the development of next-generation immunotherapies aimed at reprogramming the tumor microenvironment and overcoming resistance.

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