Co-culture

Tumor–immune cell co-culture models are a type of in vitro research model in which tumor cells and immune cells are cultured together to simulate the interactions between tumor cells and immune cells within the tumor microenvironment (TME). Compared with tumor cells cultured alone, co-culture systems introduce immune cell-mediated processes, including recognition, activation, cytotoxicity, and immune suppression. Therefore, these models can be used to study tumor immune escape, immune cell functions, and responses to immunotherapy.

Tumor cells + T cells

The co-culture of tumor cells and T cells is one of the most widely used tumor–immune co-culture models and is commonly applied to study T-cell recognition, activation, and killing of tumor cells. When tumor-reactive T cells are co-cultured with tumor cells, T cells can recognize tumor-associated antigens and induce cytokine production, degranulation, and cytotoxic responses, ultimately leading to tumor cell death.

Researchers can further co-culture T cells with patient-derived tumoroids or tumor organoids, allowing T cells to interact with 3D models that more closely preserve the characteristics of patient tumors. These models can be used to evaluate T-cell infiltration, tumor recognition, cytotoxicity, and responses to immunotherapy, and can also be applied to investigate cellular immunotherapy strategies including immune checkpoint blockade, CAR-T, and TCR-T therapies.

Tumor cells + NK cells

The co-culture model of tumor cells and natural killer cells (NK cells) is mainly used to study NK-cell-mediated tumor recognition and non-antigen-specific cytotoxicity. NK cells can recognize certain abnormal or stressed tumor cells without classical antigen-specific recognition and induce tumor cell death through the release of perforin, granzymes, and other effector molecules.

In co-culture systems, NK-cell function can be evaluated by detecting activation markers, degranulation responses, cytokine production, and tumor cell survival rates. Furthermore, combining NK cells with 3D tumor spheroids or tumoroids enables the investigation of NK-cell migration and infiltration into tumor structures, as well as the suppressive effects of the tumor microenvironment on NK-cell function.

Therefore, Tumor–NK cell co-culture models are widely used for studying NK-cell-based immunotherapy, antibody-dependent cellular cytotoxicity (ADCC), tumor immune escape, and regulation of NK-cell function.

Tumor cells + Macrophages

The co-culture model of tumor cells and macrophages is mainly used to simulate interactions between tumor cells and tumor-associated macrophages (TAMs). Tumor cells can regulate macrophage recruitment and functional states by secreting cytokines, chemokines, and other signaling molecules. In turn, macrophages can influence tumor cell proliferation, invasion, and therapeutic responses through the secretion of growth factors, inflammatory mediators, and immune regulatory molecules.

In vitro co-culture systems allow researchers to investigate tumor cell-induced macrophage phenotype changes. Macrophage states can be analyzed using markers such as CD68, CD163, CD206, and MHC-II, together with cytokine expression analysis. Previous studies have established co-culture models using NSCLC tumor cells and macrophages to investigate how different tumor cells regulate macrophage phenotypes and functions.

By further incorporating macrophages into 3D tumor spheroids or tumoroids, researchers can study macrophage recruitment and infiltration into tumor tissues, as well as their effects on tumor growth, invasion, and drug responses. Recently, tumoroid–macrophage co-culture models have also been used to simulate more complex tumor microenvironments and evaluate tumor–immune interactions and therapeutic responses.

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