Generation of monocyte-derived tumor-associated macrophages using tumor-conditioned media provides a novel method to study tumor-associated macrophages in vitro

Autor: Kallie Jiang, John Harrison Howard, Amanda Campbell, William E. Carson, Thomas A. Mace, Megan C. Duggan, Robert A. Baiocchi, Susheela Tridandapani, Luke Scarberry, Brooke Benner, Gabriella Lapurga, Emily Smith, Lorena P. Suarez-Kelly, Jonathan P. Butchar
Rok vydání: 2018
Předmět:
0301 basic medicine
Cancer Research
medicine.medical_treatment
Immunology
CCL2
lcsh:RC254-282
03 medical and health sciences
0302 clinical medicine
stomatognathic system
In vivo
Cell Line
Tumor
medicine
Tumor Microenvironment
Immunology and Allergy
Humans
skin and connective tissue diseases
Cancer
Pharmacology
Tumor microenvironment
Chemistry
Monocyte
Macrophages
Tumor-associated macrophages
In vitro generation
Cell Differentiation
Immunotherapy
lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens
In vitro
3. Good health
030104 developmental biology
Cytokine
medicine.anatomical_structure
Cell Transformation
Neoplastic
Oncology
Cell culture
030220 oncology & carcinogenesis
Culture Media
Conditioned
Cancer research
Molecular Medicine
hormones
hormone substitutes
and hormone antagonists
Research Article
Zdroj: Journal for Immunotherapy of Cancer
Journal for ImmunoTherapy of Cancer, Vol 7, Iss 1, Pp 1-14 (2019)
ISSN: 2051-1426
Popis: Background Tumor-associated macrophages (TAM) are expanded and exhibit tumor-promoting properties within the tumor microenvironment. Current methods to study TAM have not been replicated across cancer types and often do not include exogenous growth factors from the tumor, a key factor in TAM differentiation in vivo. Methods In this study, an in vitro method to generate monocyte- derived TAM using tumor- conditioned media (TCM) and a cytokine cocktail containing IL-4, IL-10, and M-CSF was utilized to study the phenotype, morphology, and function of TAM across multiple cancer types. TCM was generated from two breast cancer cell lines and an Epstein-Barr virus-positive lymphoma cell line. The properties of in vitro generated TAM were compared to in vitro generated M1 and M2- like macrophages and TAM isolated from patients with cancer. Results TAM generated in this fashion displayed an increase in CD163/CD206 co-expression compared to M2- like macrophages (87 and 36%, respectively). TAM generated in vitro exhibited increased transcript levels of the functional markers IL-6, IL-10, CCL2, c-Myc, iNOS, and arginase compared to in vitro generated M2-like macrophages. Functionally, in vitro generated TAM inhibited the proliferation of T cells (47% decrease from M1-like macrophages) and the production of IFN-γ by natural killer cells was inhibited (44%) when co-cultured with TAM. Furthermore, in vitro generated TAM secreted soluble factors that promote the growth and survival of tumor cells. Conclusions Limited access to patient TAM highlights the need for methods to generate TAM in vitro. Our data confirm that monocyte-derived TAM can be generated reliably using TCM plus the cytokine cocktail of IL-4, IL-10, and M-CSF. Given the ability of TAM to inhibit immune cell function, continued study of methods to deplete or deactivate TAM in the setting of cancer are warranted. Electronic supplementary material The online version of this article (10.1186/s40425-019-0622-0) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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