Immunocompatibility Group, Department of Pharmacology and Therapeutics, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool L7 3NY, UK.
Centre of Excellence for Long-Acting Therapeutics (CELT), Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool L7 8TX, UK.
Int J Mol Sci. 2024 May 17;25(10):5491. doi: 10.3390/ijms25105491.
Macrophages are well known for their involvement in the biocompatibility, as well as biodistribution, of nano(bio)materials. Although there are a number of rodent cell lines, they may not fully recapitulate primary cell responses, particularly those of human cells. Isolation of tissue-resident macrophages from humans is difficult and may result in insufficient cells with which to determine the possible interaction with nano(bio)materials. Isolation of primary human monocytes and differentiation to monocyte-derived macrophages may provide a useful tool with which to further study these interactions. To that end, we developed a standard operating procedure for this differentiation, as part of the Regulatory Science Framework for Nano(bio)material-based Medical Products and Devices (REFINE) project, and used it to measure the secretion of bioactive molecules from M1 and M2 differentiated monocytes in response to model nano(bio)materials, following an initial assessment of pyrogenic contamination, which may confound potential observations. The SOP was deployed in two partner institutions with broadly similar results. The work presented here shows the utility of this assay but highlights the relevance of donor variability in responses to nano(bio)materials. Whilst donor variability can provide some logistical challenges to the application of such assays, this variability is much closer to the heterogeneous cells that are present in vivo, compared to homogeneous non-human cell lines.
巨噬细胞在纳米(生物)材料的生物相容性和生物分布中起着重要作用。尽管有许多啮齿动物细胞系,但它们可能无法完全重现原代细胞的反应,尤其是人类细胞的反应。从人体中分离组织驻留巨噬细胞很困难,并且可能导致获得的细胞数量不足,无法确定与纳米(生物)材料的可能相互作用。分离原代人单核细胞并分化为单核细胞来源的巨噬细胞可能为进一步研究这些相互作用提供有用的工具。为此,我们开发了一种标准操作规程来进行这种分化,这是基于监管科学框架的纳米(生物)材料医疗产品和设备(REFINE)项目的一部分,并在对热源污染进行初步评估后,使用该方法来测量 M1 和 M2 分化的单核细胞对模型纳米(生物)材料的生物活性分子的分泌,这可能会干扰潜在的观察结果。该 SOP 在两家具有广泛相似结果的合作机构中得到了部署。这里呈现的工作展示了该测定的实用性,但强调了对纳米(生物)材料的反应中供体变异性的相关性。虽然供体变异性可能会对这种测定的应用带来一些实际挑战,但与非人类细胞系相比,这种变异性更接近体内存在的异质细胞。