Isolation of mouse and human tumor-associated macrophages

Luca Cassetta, Roy Noy, Agnieszka Swierczak, Gaël Sugano, Harriet O. Smith, Lisa Wiechmann, Jeffrey W. Pollard

Research output: Contribution to journalArticle

15 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)211-229
Number of pages19
JournalAdvances in Experimental Medicine and Biology
Volume899
DOIs
StatePublished - 2016
Externally publishedYes

Fingerprint

Macrophages
Tumors
Neoplasms
Cells
Tissue
Network protocols
Flow cytometry
Immune system
Tumor Microenvironment
Complex networks
Cell Separation
Transcriptome
Immunosuppression
Immune System
Flow Cytometry

Keywords

  • Breast cancer
  • Flow cytometry
  • Human
  • Macrophages
  • Mouse
  • TAM
  • Tissue
  • Tumor-associated macrophages

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Cassetta, L., Noy, R., Swierczak, A., Sugano, G., Smith, H. O., Wiechmann, L., & Pollard, J. W. (2016). Isolation of mouse and human tumor-associated macrophages. Advances in Experimental Medicine and Biology, 899, 211-229. https://doi.org/10.1007/978-3-319-26666-4_12

Isolation of mouse and human tumor-associated macrophages. / Cassetta, Luca; Noy, Roy; Swierczak, Agnieszka; Sugano, Gaël; Smith, Harriet O.; Wiechmann, Lisa; Pollard, Jeffrey W.

In: Advances in Experimental Medicine and Biology, Vol. 899, 2016, p. 211-229.

Research output: Contribution to journalArticle

Cassetta, L, Noy, R, Swierczak, A, Sugano, G, Smith, HO, Wiechmann, L & Pollard, JW 2016, 'Isolation of mouse and human tumor-associated macrophages', Advances in Experimental Medicine and Biology, vol. 899, pp. 211-229. https://doi.org/10.1007/978-3-319-26666-4_12
Cassetta, Luca ; Noy, Roy ; Swierczak, Agnieszka ; Sugano, Gaël ; Smith, Harriet O. ; Wiechmann, Lisa ; Pollard, Jeffrey W. / Isolation of mouse and human tumor-associated macrophages. In: Advances in Experimental Medicine and Biology. 2016 ; Vol. 899. pp. 211-229.
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