从原位 4T1 乳腺癌肿瘤中分离和分选小鼠肿瘤相关巨噬细胞（TAMs）亚群。

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

机构信息

Laboratory of Tumor Inflammation and Angiogenesis, Center for Cancer Biology, VIB, Leuven B3000, Belgium.

Laboratory of Tumor Inflammation and Angiogenesis, Center for Cancer Biology, Department of Oncology, KU Leuven, Leuven B3000, Belgium.

出版信息

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

DOI:10.1016/j.xpro.2021.100481

PMID:33982015

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8082159/

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).

摘要

肿瘤相关巨噬细胞（TAMs）在肿瘤内定位、表面标志物表达和分子特性方面具有高度异质性。本方案描述了从小鼠原位 4T1 乳腺癌肿瘤中分离和消化小鼠乳腺癌样本以及通过荧光激活细胞分选（FACS）分离 TAMs 的完整过程。这包括分离 PoEMs（表达 podoplanin 的巨噬细胞）和非 PoEMs（podoplanin 阴性巨噬细胞）的步骤。我们的 FACS 分离方法也可用于其他浸润 TAM 的肿瘤类型。有关使用和执行本方案的完整详细信息，请参见 Bieniasz-Krzywiec 等人。（2019 年）。