Abstract
Objectives
To identify transcription markers that uniquely determine specific Chinese hamster ovary (CHO) cell lines and can be used for the identification of cell lines in the process of biopharmaceutical cell-line development.
Results
Five CHO cell lines with different origins were extensively characterised at the transcriptomic level and the results were compared to their karyotype characterisation. The analysed cell lines differ in their karyotype but, due to the genome instability observed during parental and recombinant cell-line establishment, karyotyping is not the preferred method for accurate identification of the various CHO cell lines. Marker genes unique to a specific cell line were identified by microarrays, and their expression was validated by reverse-transcription quantitative real-time PCR. The analysed cell lines can be differentiated by the presence/absence of detectable marker gene expression. Additionally, the similarity of the transcriptional profiles is dependent on cell-line history but independent of the manipulation steps involved in the recombinant cell-line development process.
Conclusions
Certain transcripts can be used as markers for the identification of a CHO cell line undergoing recombinant development and thus represent a powerful tool for ensuring the maintenance of high quality standards.
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Acknowledgments
We would like to thank Dr. Holger Laux and his team for their hospitality and assistance during our visit to their laboratory. We are grateful to Katja Glinšek for her technical assistance in the karyotype analysis.
Supporting information
Supplementary Table 1—List of genes and their descriptions with the primer and probe sequences.
Supplementary Table 2—Summary of the RT-qPCR results for the genes indentified with unique expression for an individual cell line, from the data given in Fig. 5.
Supplementary Table 3—Quantitative real-time PCR analysis performed on genomic DNA isolated from the parental cell line samples.
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Blas, M., Francky, A., Jamnikar, U. et al. Transcriptomic variation between different Chinese hamster ovary cell lines. Biotechnol Lett 37, 1737–1745 (2015). https://doi.org/10.1007/s10529-015-1850-0
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DOI: https://doi.org/10.1007/s10529-015-1850-0