Pamies David, Hartung Thomas
Center for Alternatives to Animal Testing (CAAT), Johns Hopkins Bloomberg School of Public Health , Baltimore, Maryland 21205, United States.
CAAT-Europe, University of Konstanz , 78464 Konstanz, Germany.
Chem Res Toxicol. 2017 Jan 17;30(1):43-52. doi: 10.1021/acs.chemrestox.6b00269. Epub 2016 Dec 5.
There is no good science in bad models. Cell culture is especially prone to artifacts. A number of novel cell culture technologies have become more broadly available in the 21st century, which allow overcoming limitations of traditional culture and are more physiologically relevant. These include the use of stem-cell derived human cells, cocultures of different cell types, scaffolds and extracellular matrices, perfusion platforms (such as microfluidics), 3D culture, organ-on-chip technologies, tissue architecture, and organ functionality. The physiological relevance of such models is further enhanced by the measurement of biomarkers (e.g., key events of pathways), organ specific functionality, and more comprehensive assessment cell responses by high-content methods. These approaches are still rarely combined to create microphysiological systems. The complexity of the combination of these technologies can generate results closer to the in vivo situation but increases the number of parameters to control, bringing some new challenges. In fact, we do not argue that all cell culture needs to be that sophisticated. The efforts taken are determined by the purpose of our experiments and tests. If only a very specific molecular target to cell response is of interest, a very simple model, which reflects this, might be much more suited to allow standardization and high-throughput. However, the less defined the end point of interest and cellular response are, the better we should approximate organ- or tissue-like culture conditions to make physiological responses more probable. Besides these technologic advances, important progress in the quality assurance and reporting on cell cultures as well as the validation of cellular test systems brings the utility of cell cultures to a new level. The advancement and broader implementation of Good Cell Culture Practice (GCCP) is key here. In toxicology, this is a major prerequisite for meaningful and reliable results, ultimately supporting risk assessment and product development decisions.
糟糕的模型产生不了好的科学成果。细胞培养特别容易产生假象。21世纪,一些新型细胞培养技术已更广泛地可得,这些技术能够克服传统培养的局限性,且在生理上更具相关性。这些技术包括使用干细胞衍生的人类细胞、不同细胞类型的共培养、支架和细胞外基质、灌注平台(如微流控技术)、3D培养、芯片器官技术、组织结构和器官功能。通过生物标志物(如信号通路的关键事件)的测量、器官特异性功能以及通过高内涵方法对细胞反应进行更全面的评估,此类模型在生理上的相关性进一步增强。这些方法仍很少结合起来创建微生理系统。这些技术组合的复杂性能够产生更接近体内情况的结果,但会增加需要控制的参数数量,带来一些新的挑战。事实上,我们并不是说所有细胞培养都需要如此复杂。所付出的努力取决于我们实验和测试的目的。如果只对细胞反应的一个非常特定的分子靶点感兴趣,一个能反映这一点的非常简单的模型可能更适合实现标准化和高通量。然而,感兴趣的终点和细胞反应定义得越不明确,我们就越应接近器官或组织样的培养条件,以使生理反应更有可能发生。除了这些技术进步外,细胞培养质量保证和报告以及细胞测试系统验证方面的重要进展将细胞培养的效用提升到了一个新水平。良好细胞培养规范(GCCP)的推进和更广泛实施是关键所在。在毒理学中,这是获得有意义且可靠结果的主要前提条件,最终支持风险评估和产品开发决策。