In vitro cell culture experiments are highly important techniques to accelerate drug discovery, conduct safety testing and reduce the need for animal studies. Therefore, automatization may help to enhance the technical precision, reduce external (including operator's) influence on the data and thus improve reliability. Prior to application in scientific studies, validation of automated systems is absolutely necessary. In this study we present the validation of two combined automated pipetting systems to conduct toxicity studies in HaCaT cells consisting of cell seeding, noxious agent exposure and several assays to assess cell survival, apoptosis and interleukin production. After initial validation of pipetting accuracy, we compared homogeneity after automated seeding to plates seeded by expert laboratory technicians. Moreover, automated dispensing of a potentially unstable noxious agent was analyzed in terms of speed and consistency. We found a 2% technical imprecision for the cell survival assay and 4.5-6% for the other assays, bioluminescent and ELISA techniques. Thus, we could demonstrate the excellent technical precision of our assays. In a final step, we found that intraday variations, though acceptable, were much larger than technical variations and had to assume an intraday biological variability between different wells of the same experimental group.

中文翻译：

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验证用于硫芥毒理学中细胞培养接种、暴露和生物分析测定的自动移液系统

体外细胞培养实验是加速药物发现、进行安全测试和减少动物研究需求的非常重要的技术。因此，自动化有助于提高技术精度，减少外部（包括操作员）对数据的影响，从而提高可靠性。在应用于科学研究之前，自动化系统的验证是绝对必要的。在这项研究中，我们展示了两个组合自动移液系统的验证，以在 HaCaT 细胞中进行毒性研究，包括细胞接种、有毒物质暴露和几种评估细胞存活、细胞凋亡和白细胞介素产生的测定。在对移液准确性进行初步验证后，我们将自动接种后的均匀性与专家实验室技术人员接种的平板进行了比较。而且，在速度和一致性方面分析了潜在不稳定有害物质的自动分配。我们发现细胞存活分析的技术不精确度为 2%，其他分析、生物发光和 ELISA 技术的技术不精确度为 4.5-6%。因此，我们可以证明我们的检测具有出色的技术精度。在最后一步，我们发现日内变化虽然可以接受，但比技术变化大得多，并且必须假设同一实验组不同井之间的日内生物变异性。