Isolation of Mouse and Human Tumor-Associated Macrophages.
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.
MRC Centre for Reproductive Health, Queen's Medical Research Institute, The University of Edinburgh, Edinburgh, EH16 4TJ, UK.
Luca
L
Cassetta
Department of Developmental and Molecular Biology, New York, NY, 10461, USA.
Roy
R
Noy
MRC Centre for Reproductive Health, Queen's Medical Research Institute, The University of Edinburgh, Edinburgh, EH16 4TJ, UK.
Agnieszka
A
Swierczak
MRC Centre for Reproductive Health, Queen's Medical Research Institute, The University of Edinburgh, Edinburgh, EH16 4TJ, UK.
Gaël
G
Sugano
Department of Obstetrics and Gynecology and Women's Health, New York, NY, 10461, USA.
Harriet
H
Smith
Department of Surgery, Albert Einstein College of Medicine, New York, NY, 10461, USA.
Lisa
L
Wiechmann
MRC Centre for Reproductive Health, Queen's Medical Research Institute, The University of Edinburgh, Edinburgh, EH16 4TJ, UK. Jeff.Pollard@ed.ac.uk.|Department of Developmental and Molecular Biology, New York, NY, 10461, USA. Jeff.Pollard@ed.ac.uk.
Jeffrey W
JW
Pollard
D000818:Animals; D001943:Breast Neoplasms / Q000473:pathology; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005434:Flow Cytometry; D006461:Hemolysis; D006801:Humans; D008264:Macrophages / Q000473:pathology*; D051379:Mice; D009369:Neoplasms / Q000473:pathology*
Breast cancer; Flow cytometry; Human; Macrophages; Mouse; TAM; Tissue; Tumor-associated macrophages