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Deciphering the limitations of immortalized hepatocyte cell lines for the study of liver cis-regulatory elements

Bellesis, A., Li, X., Moore-Frederick, D., Xu, D., Delbridge, K., Ma, J., Vaccaro, G., Edward, B. A. A., Kellogg, M., Creeger, Y., et al.
10.64898/2026.06.05.730479 · was preprinted
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Abstract

Immortalized cell lines are widely used in biological research despite their known differences from their tissues and cell types of origin. Such cell lines are especially popular for testing hypotheses regarding the activity of cis-regulatory elements (CREs) that regulate gene expression. Previous investigations of blood and skin cell lines revealed many differences between the transcriptional regulatory networks of the cell lines and the associated primary cells. Similar comparisons for other tissues have been limited. Here, we used ATAC-seq to profile CREs in four immortalized liver cell lines and found many differences between each cell lines CREs and primary liver tissue, including differences in the transcription factors that are likely to bind them and differences in the genes that they are likely to regulate. Modifying cell culture conditions based on recommendations in the literature did not improve the similarity with primary liver tissue. Our results suggest that differences between the transcriptional regulatory networks in cell lines and primary tissue should be considered when designing and interpreting cell line experiments.

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  • bioRxiv Genomics @biorxiv-genomic.bsky.social · 6972 followers neutral

    Deciphering the limitations of immortalized hepatocyte cell lines for the study of liver cis-regulatory elements https://www.biorxiv.org/content/10.64898/2026.06.05.730479v1

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  • bioRxivpreprint @biorxivpreprint.bsky.social · 8892 followers neutral

    Deciphering the limitations of immortalized hepatocyte cell lines for the study of liver cis-regulatory elements https://www.biorxiv.org/content/10.64898/2026.06.05.730479v1

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