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Isolation of Mouse and Human Tumor-Associated Macrophages

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Tumor Microenvironment

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 899))

Abstract

The tumor microenvironment is a complex network of cells that support tumor progression and malignancy. It has been demonstrated that tumor cells can educate the immune system to promote a tumor-friendly environment. Among all these immune cells, tumor-associated macrophages (TAMs) are well represented and their presence in mouse models has been shown to promote tumor progression and metastasis. These effects are through the stimulation of angiogenesis, enhancement of tumor cell invasion and intravasation, immunosuppression, and at the metastatic site tumor cell extravasation and growth. However, the precise mechanisms are not fully understood. Furthermore there is limited information on TAMs derived from human cancers. For this reason it is important to be able to extract TAMs from tumors in order to compare their phenotypes, functions, and transcriptomes with normal resident tissue macrophages. Isolation of these cells is challenging due to the lack of markers and standardized protocols. Here we show an optimized protocol for the efficient isolation and extraction of resident macrophages and TAMs from human and mouse tissues by using multicolor flow cytometry. These protocols allow for the extraction of thousands of macrophages in less than 5 h from tissues as small as half a gram. The isolated macrophages can then be used for both “omics” and in vitro studies.

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Acknowledgments

We would like to thank Dr Lisa Coussens, OHSU, for valuable discussions. This work was supported by Wellcome Trust (101067/Z/13/Z) and NIH R01CA172451 to JWP and Department of Defense (DOD W81XWH-111-0702) to Dr Lisa Coussens and JWP.

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Authors and Affiliations

  1. MRC Centre for Reproductive Health, Queen’s Medical Research Institute, The University of Edinburgh, Edinburgh, EH16 4TJ, UK

    Luca Cassetta, Agnieszka Swierczak, Gaël Sugano & Jeffrey W. Pollard

  2. Department of Developmental and Molecular Biology, New York, NY, 10461, USA

    Roy Noy & Jeffrey W. Pollard

  3. Department of Obstetrics and Gynecology and Women’s Health, New York, NY, 10461, USA

    Harriet Smith

  4. Department of Surgery, Albert Einstein College of Medicine, New York, NY, 10461, USA

    Lisa Wiechmann

Authors
  1. Luca Cassetta
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  2. Roy Noy
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  3. Agnieszka Swierczak
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  4. Gaël Sugano
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  5. Harriet Smith
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  6. Lisa Wiechmann
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  7. Jeffrey W. Pollard
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Corresponding author

Correspondence to Jeffrey W. Pollard .

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Editors and Affiliations

  1. Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Constantinos Koumenis

  2. Cell & Developmental Biology, Oregon Health & Science University, Portland, Oregon, USA

    Lisa M. Coussens

  3. Department of Radiation Oncology, Stanford University School of Medic, Stanford, California, USA

    Amato Giaccia

  4. Department of Oncology, University of Oxford, Oxford, Oxfordshire, United Kingdom

    Ester Hammond

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© 2016 Springer International Publishing Switzerland

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Cassetta, L. et al. (2016). Isolation of Mouse and Human Tumor-Associated Macrophages. In: Koumenis, C., Coussens, L., Giaccia, A., Hammond, E. (eds) Tumor Microenvironment. Advances in Experimental Medicine and Biology, vol 899. Springer, Cham. https://doi.org/10.1007/978-3-319-26666-4_12

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