Illuminating Clonal Dynamics: Development and Use of a High-Throughput Cellular Barcoding System to Track Clonal Evolution
Porter, Shaina N. 1983-
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It is increasingly recognized that tracking the clonal dynamics of large populations is important in understanding aspects of cancer and stem cell biology. Attempts to track the contributions of individual cells to the clonality of large homogenous populations, however, have been constrained by limitations in sensitivity and complexity. We have created an efficient and high throughput method to overcome these limitations by harnessing the power of viral marking and next generation sequencing technology, allowing us to track the clonal contributions of many thousands of cells with minimal perturbations to the population as a whole. We have applied this system to several of the most commonly used cell lines in biological research in order to validate this system and gain valuable biological insight into these ubiquitous research tools. Cell lines, often continuously passaged for years, are often assumed to be clonal, the most fit and stable clone having been selected during establishment and early passages. However, our results show that ongoing genomic and/or epigenomic instability within these cells leads to proliferative instability, resulting in the divergence of clones from one another and revealing unexpected clonal dynamics and rapid clonal evolution. These results have profound implications for the experimental use of cell lines, as well as broad applications in the fields of stem cells and cancer biology, among others.