Isolation of mouse mammary carcinoma-derived macrophages and cancer cells for co-culture assays

Sun, Lijuan, Han, Xiao, Egeblad, Mikala (December 2022) Isolation of mouse mammary carcinoma-derived macrophages and cancer cells for co-culture assays. STAR Protocols, 3 (4). p. 101833. ISSN 2666-1667 (In Press)

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Abstract

We recently established an in vitro co-culture system in which monophosphoryl lipid A + interferon-γ (MPLA+IFNγ)-treated tumor-associated macrophages (TAMs) killed cancer cells. Here, we describe a step-by-step protocol for isolating TAMs and cancer cells from mouse primary mammary carcinomas, the setup of the co-culture system, and the image acquisition approach. The technical difficulties in the co-culture assay involve isolating pure TAMs and cancer cells from the same tumor and staining them with different dyes to track the macrophages' tumoricidal activity. For complete details on the use and execution of this protocol, please refer to Sun et al.(2021).

Item Type:	Paper
Subjects:	diseases & disorders > cancer Investigative techniques and equipment organism description > animal Investigative techniques and equipment > assays diseases & disorders > cancer > cancer types > breast cancer Investigative techniques and equipment > cell culture organs, tissues, organelles, cell types and functions > cell types and functions > cell types organs, tissues, organelles, cell types and functions > cell types and functions > cell types organs, tissues, organelles, cell types and functions > cell types and functions > cell types organs, tissues, organelles, cell types and functions > cell types and functions organs, tissues, organelles, cell types and functions > cell types and functions > cell types > macrophages organs, tissues, organelles, cell types and functions > cell types and functions > cell types > macrophages organs, tissues, organelles, cell types and functions > cell types and functions > cell types > macrophages organism description > animal > mammal organism description > animal > mammal > rodent > mouse organs, tissues, organelles, cell types and functions organism description > animal > mammal > rodent diseases & disorders > cancer > cancer types
CSHL Authors:	Sun, Lijuan Han, Xiao Egeblad, Mikala
Communities:	CSHL labs > Egeblad lab CSHL Cancer Center Program > Cellular Communication in Cancer Program
SWORD Depositor:	CSHL Elements
Depositing User:	CSHL Elements
Date:	December 2022
Date Deposited:	22 Nov 2022 18:00
Last Modified:	09 Feb 2024 20:39
PMCID:	PMC9664409
URI:	https://repository.cshl.edu/id/eprint/40760

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