Laboratory of Molecular OncoSignalling, IMol Polish Academy of Sciences, Flisa 6, Warsaw, 02-247, Poland.
J Mammary Gland Biol Neoplasia. 2024 Feb 10;29(1):4. doi: 10.1007/s10911-024-09556-2.
Tumor mass comprises not only cancer cells but also heterogeneous populations of immune and stromal cells, along with the components of the extracellular matrix, collectively called the tumor microenvironment (TME). This diverse population of cells can communicate with each other, which can positively or negatively affect tumor growth and progression to malignancy. The most common type of immune cells in the TME are macrophages. Macrophages continuously differentiate into a broad landscape of tumor-associated macrophages (TAMs) in response to numerous signals from the TME, which makes studies on TAMs quite challenging. Therefore, implementing reliable protocols is a milestone for drawing consistent conclusions about the interactions between cancer cells and TAMs. Here, we provide the details for the polarization of a human leukemia monocytic cell line, THP-1, into M0, M1 and M2 macrophages. We also present a step-by-step protocol for a transwell co-culture using a human breast cancer cell line, HCC1806, and THP-1-derived macrophages. Finally, we describe the colony formation and migration assays performed on the breast cancer cells after the co-culture with macrophages to measure the influence of macrophages on the oncogenic features of cancer cells. In summary, our co-culture-based protocols can be a valuable resource for investigating the interactions between macrophages and cancer cells.
肿瘤块不仅包含癌细胞，还包含异质的免疫细胞和基质细胞群体，以及细胞外基质的成分，统称为肿瘤微环境（TME）。这些不同的细胞群体可以相互交流，这可能会对肿瘤的生长和恶性转化产生积极或消极的影响。TME 中最常见的免疫细胞类型是巨噬细胞。巨噬细胞会根据 TME 中的众多信号不断分化为广泛的肿瘤相关巨噬细胞（TAMs），这使得 TAMs 的研究极具挑战性。因此，实施可靠的方案是得出关于癌细胞和 TAMs 之间相互作用的一致结论的一个里程碑。在这里，我们提供了将人白血病单核细胞系 THP-1 极化为人 M0、M1 和 M2 巨噬细胞的详细信息。我们还介绍了使用人乳腺癌细胞系 HCC1806 和 THP-1 衍生的巨噬细胞进行 Transwell 共培养的分步方案。最后，我们描述了在与巨噬细胞共培养后对乳腺癌细胞进行集落形成和迁移分析的实验，以测量巨噬细胞对癌细胞致癌特征的影响。总之，我们基于共培养的方案可以成为研究巨噬细胞和癌细胞之间相互作用的有价值的资源。
