In Vitro Differentiation of Tumor-Associated Macrophages from Monocyte Precursors with Modified Melanoma-Conditioned Medium

Methods Mol Biol. 2021:2265:119-128. doi: 10.1007/978-1-0716-1205-7_9.

Abstract

Tumor-associated macrophages (TAMs) are one of most important components of the tumor microenvironment. Although many assays have been developed to differentiate monocytes into macrophages (Mϕ) for studying the biology of TAMs in vitro, little is known whether the macrophages induced by these approaches can recapitulate the biology of TAMs present in the tumor microenvironment. We have developed a novel assay to differentiate human monocytes into TAMs using modified melanoma-conditioned medium, which is derived from the concentrated tumor cell culture medium. Characterization of these modified melanoma-conditioned medium-induced macrophages (MCMI-Mϕ) by multiple flow cytometry, Luminex, microarray, and immunohistochemistry analyses indicates that MCMI-Mϕ are phenotypically and functionally highly similar to the TAMs present in the tumor microenvironment.

Keywords: Macrophages; Modified melanoma-conditioned medium; Monocyte; Tumor cells.

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Differentiation / drug effects*
  • Cell Line, Tumor
  • Culture Media, Conditioned / pharmacology*
  • Humans
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / drug effects
  • Macrophages / cytology*
  • Melanoma / metabolism*
  • Melanoma / pathology
  • Monocytes / cytology
  • Monocytes / drug effects
  • Tumor Microenvironment
  • Tumor-Associated Macrophages

Substances

  • Culture Media, Conditioned