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首页> 外文期刊>Journal of Leukocyte Biology: An Official Publication of the Reticuloendothelial Society >Technical Advance: An optimized disaggregation method for human lung tumors that preserves the phenotype and function of the immune cells
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Technical Advance: An optimized disaggregation method for human lung tumors that preserves the phenotype and function of the immune cells

机译：技术进步：一种优化的人肺肿瘤分解方法，可保留免疫细胞的表型和功能

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摘要

Careful preparation of human tissues is the cornerstone of obtaining accurate data in immunologic studies. Despite the essential importance of tissue processing in tumor immunology and clinical medicine, current methods of tissue disaggregation have not been rigorously tested for data fidelity. Thus, we critically evaluated the current techniques available in the literature that are used to prepare human lung tumors for immunologic studies. We discovered that these approaches are successful at digesting cellular attachments and ECMs; however, these methods frequently alter the immune cell composition and/or expression of surface molecules. We thus developed a novel approach to prepare human lung tumors for immunologic studies by combining gentle mechanical manipulation with an optimized cocktail of enzymes used at low doses. This enzymatic digestion cocktail optimized cell yield and cell viability, retrieved all major tumor-associated cell populations, and maintained the expression of cell-surface markers for lineage definition and in vivo effector functions. To our knowledge, we present the first rigorously tested disaggregation method designed for human lung tumors.
机译：精心准备人体组织是获得免疫学研究准确数据的基石。尽管组织加工在肿瘤免疫学和临床医学中至关重要，但目前尚未对组织分解的方法进行严格的数据保真度测试。因此，我们严格评估了文献中可用于制备用于免疫学研究的人肺肿瘤的当前技术。我们发现这些方法在消化细胞附着和ECM方面是成功的。然而，这些方法经常改变免疫细胞的组成和/或表面分子的表达。因此，我们通过结合轻柔的机械操作和优化的低剂量酶混合物，开发了一种用于免疫学研究的人肺肿瘤制备新方法。这种酶消化混合物优化了细胞产量和细胞活力，回收了所有主要的肿瘤相关细胞群，并维持了细胞表面标志物的表达，以用于谱系定义和体内效应子功能。据我们所知，我们提出了第一种经过严格测试的针对人肺肿瘤的分解方法。

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