Petiti Jessica, Caria Sabrina, Revel Laura, Pegoraro Mattia, Divieto Carla
Division of Advanced Materials Metrology and Life Sciences, Istituto Nazionale di Ricerca Metrologica (INRIM), 10135 Turin, Italy.
Department of Chemistry, University of Turin, 10125 Turin, Italy.
Cells. 2024 Nov 26;13(23):1959. doi: 10.3390/cells13231959.
The A549 cell line has become a cornerstone in biomedical research, particularly in cancer studies and serves as a critical tool in cytotoxicity studies and drug screening where it is used to evaluate the impact of pharmaceutical compounds on cellular viability. One of the most widely adopted methods for viability assessment, which is also used in evaluating drug cytotoxicity, is the resazurin-based assay. This assay exploits the ability of living cells to convert resazurin into fluorescent resorufin, providing a reliable indicator of metabolic activity. By measuring this conversion, cell viability can be estimated. Resazurin assay is extensively used for evaluating cytotoxic effects on various cell lines, including A549 cells, thereby bridging the gap between in vitro experimentation and drug development. However, frequent data inconsistencies in pre-clinical drug screening highlight the critical need for standardization to ensure reliability and reproducibility. This manuscript addresses these challenges by describing the optimization of resazurin-based viability assays for A549 cells in both 2D cultures and 3D fibrin gel models. By optimizing this test, the study aims to enhance the reliability of cytotoxicity results and introduces a new standard operating procedure, thus providing consistent results with minimal measurement uncertainty. This standardization is crucial for advancing drug screening and ensuring robust research findings.
A549细胞系已成为生物医学研究的基石，尤其是在癌症研究中，并作为细胞毒性研究和药物筛选的关键工具，用于评估药物化合物对细胞活力的影响。基于刃天青的检测法是评估细胞活力最广泛采用的方法之一，也用于评估药物细胞毒性。该检测法利用活细胞将刃天青转化为荧光试卤灵的能力，提供代谢活性的可靠指标。通过测量这种转化，可以估计细胞活力。刃天青检测法广泛用于评估对包括A549细胞在内的各种细胞系的细胞毒性作用，从而弥合体外实验与药物开发之间的差距。然而，临床前药物筛选中频繁的数据不一致突出了标准化的迫切需求，以确保可靠性和可重复性。本手稿通过描述在二维培养和三维纤维蛋白凝胶模型中对A549细胞基于刃天青的活力检测法的优化来解决这些挑战。通过优化该检测，本研究旨在提高细胞毒性结果的可靠性，并引入新的标准操作规程，从而以最小的测量不确定性提供一致的结果。这种标准化对于推进药物筛选和确保可靠的研究结果至关重要。
