Isolation and separation of murine tumor-associated macrophages (TAMs) subpopulations from orthotopic 4T1 breast tumors

STAR Protoc. 2021 Apr 17;2(2):100481. doi: 10.1016/j.xpro.2021.100481. eCollection 2021 Jun 18.

Abstract

Tumor-associated macrophages (TAMs) are highly heterogenous regarding their intratumoral localization, surface marker expression, and molecular properties. This protocol describes the complete procedure for isolation and digestion of murine breast cancer samples and fluorescence-activated cell sorting (FACS) of TAMs from murine orthotopic 4T1 breast tumors. This includes steps to separate PoEMs (podoplanin-expressing macrophages) and non-PoEMs (podoplanin-negative macrophages). Our FACS separation approach could also be used for other tumor types with TAM infiltration. For complete details on the use and execution of this protocol, please refer to Bieniasz-Krzywiec et al. (2019).

Keywords: Cancer; Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Model Organisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Breast Neoplasms* / chemistry
  • Breast Neoplasms* / metabolism
  • Breast Neoplasms* / pathology
  • Cell Culture Techniques
  • Cell Line, Tumor
  • Cells, Cultured
  • Female
  • Flow Cytometry / methods*
  • Mice
  • Tumor-Associated Macrophages / chemistry
  • Tumor-Associated Macrophages / cytology*