Department of Pharmacology, Yale University School of Medicine, New Haven, CT, USA.
Yale Cancer Biology Institute, Yale University, West Haven, CT, USA.
Nat Biotechnol. 2019 Mar;37(3):314-322. doi: 10.1038/s41587-019-0037-y. Epub 2019 Feb 18.
Reproducibility in research can be compromised by both biological and technical variation, but most of the focus is on removing the latter. Here we investigate the effects of biological variation in HeLa cell lines using a systems-wide approach. We determine the degree of molecular and phenotypic variability across 14 stock HeLa samples from 13 international laboratories. We cultured cells in uniform conditions and profiled genome-wide copy numbers, mRNAs, proteins and protein turnover rates in each cell line. We discovered substantial heterogeneity between HeLa variants, especially between lines of the CCL2 and Kyoto varieties, and observed progressive divergence within a specific cell line over 50 successive passages. Genomic variability has a complex, nonlinear effect on transcriptome, proteome and protein turnover profiles, and proteotype patterns explain the varying phenotypic response of different cell lines to Salmonella infection. These findings have implications for the interpretation and reproducibility of research results obtained from human cultured cells.
研究的可重复性可能受到生物和技术变异的影响，但大多数关注的是消除后者。在这里，我们采用系统的方法研究了 HeLa 细胞系中的生物学变异的影响。我们确定了来自 13 个国际实验室的 14 个库存 HeLa 样本的分子和表型变异程度。我们在统一的条件下培养细胞，并对每个细胞系的全基因组拷贝数、mRNA、蛋白质和蛋白质周转速率进行了分析。我们发现 HeLa 变体之间存在很大的异质性，尤其是 CCL2 和京都品系之间的差异，并且在特定细胞系中经过 50 多次连续传代后观察到逐渐的分歧。基因组的变异性对转录组、蛋白质组和蛋白质周转速率的影响是复杂的、非线性的，蛋白质组模式解释了不同细胞系对沙门氏菌感染的不同表型反应。这些发现对从人类培养细胞中获得的研究结果的解释和可重复性具有重要意义。
