Tracing production instability in a clonally derived CHO cell line using single-cell transcriptomics

A variety of mechanisms including transcriptional silencing, gene copy loss, and increased susceptibility to cellular stress have been associated with a sudden or gradual loss of monoclonal antibody (mAb) production in Chinese hamster ovary (CHO) cell lines. In this study, we utilized single-cell RNA-seq (scRNA-seq) to study a clonally derived CHO cell line that underwent production instability leading to a dramatic reduction of the levels of mAb produced. From the scRNA-seq data, we identified subclusters associated with variations in the mAb transgenes and observed that heavy chain gene expression was significantly lower than that of the light chain across the population. Using trajectory inference, the evolution of the cell line was reconstructed and was found to correlate with a reduction in heavy and light chain gene expression. Genes encoding for proteins involved in the response to oxidative stress and apoptosis were found to increase in expression as cells progressed along the trajectory. Future studies of CHO cell lines using this technology have the potential to dramatically enhance our understanding of the characteristics underpinning efficient manufacturing performance as well as product quality.

Automated summary

The study used single-cell RNA sequencing to investigate the production instability of a clonally derived CHO cell line, revealing that heavy chain gene expression was significantly lower compared to that of the light chain. The evolution of the cell line showed a reduction in heavy and light chain gene expression, while genes involved in oxidative stress and apoptosis response increased in expression as cells progressed along the trajectory. Future studies utilizing this technology have the potential to enhance understanding of efficient manufacturing performance and product quality in CHO cell lines.