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We identified widespread intercellular heteroplasmy across the CHO cell population and predicted possible phenotypic impacts. 3\/11 (27%) of the most variable mutations were only identified by scmtDNAseq, indicating greater resolution when compared to bulk cell analysis. Single\u2010cell RNAseq (scRNAseq) was also performed at the same time point and, compared to scmtDNAseq, significant differences in intercellular heteroplasmy were observed. Using an inducible mAb expression system demonstrated that short\u2010term additional biosynthetic burden of exogenous protein production had little impact on intercellular heteroplasmy. We additionally monitored bulk heteroplasmy over 38 days, reflecting the typical timespan from vial thaw to production vessel in a Biopharmaceutical upstream cell culture process. We observed minimal change in heteroplasmy, finding no evidence that a mAb\u2010producing CHO cell line develops impactful changes in heteroplasmy over that timeframe. This would suggest that for our cell line, the heteroplasmy profile established on Day 1 should be maintained throughout a full fed\u2010batch bioprocess run.<\/jats:p>","DOI":"10.1002\/biot.70079","type":"journal-article","created":{"date-parts":[[2025,9,1]],"date-time":"2025-09-01T08:33:23Z","timestamp":1756715603000},"source":"Crossref","is-referenced-by-count":1,"title":["Single\u2010Cell Mitochondrial DNA Analysis of Recombinant Chinese Hamster Ovary Cells Reveals Widespread Heteroplasmy"],"prefix":"10.1002","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0535-9106","authenticated-orcid":false,"given":"Alan","family":"Foley","sequence":"first","affiliation":[{"name":"Cell Engineering Group NIBRT  Dublin Ireland"},{"name":"School of Chemical and Bioprocess Engineering University College Dublin  Dublin 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