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Purification of Tumor-Associated Macrophages (TAM) and Tumor-Associated Dendritic Cells (TADC)   

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Original research article

A brief version of this protocol appeared in:
Cancer Research
Jan 2014

Abstract

Tumors are heterogeneous microenvironments where complex interactions take place between neoplastic cells and infiltrating inflammatory cells, such as tumor-associated macrophages (TAM) and tumor-associated dendritic cells (TADC). The relevance of tumor-infiltrating mononuclear myeloid cells is underscored by clinical studies showing a correlation between their abundance and poor prognosis (Laoui et al., 2011). These cells are able to promote tumor progression via several mechanisms, including induction of angiogenesis, remodeling of the extracellular matrix, stimulation of cancer cell proliferation and metastasis and the inhibition of adaptive immunity (Laoui et al., 2011). Moreover, mononuclear myeloid cells are characterized by plasticity and versatility in response to microenvironmental signals, resulting in different activation states, as illustrated by the presence of distinct functional TAM subsets in tumors (Movahedi et al., 2010; Laoui et al., 2014). Here, we describe a valuable isolation technique for TAM and TADC permitting their molecular and functional characterization.

Keywords: Tumor-associated macrophages, Tumor-associated dendritic cells, Tumor single cell suspension, Macrophage purification, Dendritic cell purification

Copyright: © 2014 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Laoui, D., Overmeire, E. V., Keirsse, J., Movahedi, K. and Ginderachter, J. A. (2014). Purification of Tumor-Associated Macrophages (TAM) and Tumor-Associated Dendritic Cells (TADC). Bio-protocol 4(22): e1294. DOI: 10.21769/BioProtoc.1294.
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Margarita Bartish
Karolinska Institutet
Hi!

I have a question with regards to your use of lymphoprep to purify the dissociated tumor: I am interested in collecting Ly6G+ cells from the tumor (putative granulocytic MDSCs). The lymphoprep description from the manufacturer (in my case, StemCell) states that "differences in cell density are exploited to separate granulocytes and erythrocytes from mononuclear cells", seemingly precluding its use for my purposes. But I see a Ly6G+ population in your facs plots. Does lymphoprep behave differently if it used on whole blood compared to as you do, dissociated tumor tissue? In your experience, based on the percentages of Ly6G+ you see, do you think it would be possible to extract a workable number of cells from a tumor sample? Thank you for your input!
11/23/2016 5:00:46 AM Reply