HeLa cell heterogeneity and coxsackievirus B3 cytopathic effect: Implications for inter‐laboratory reproducibility of results

Concerns over cell line identities and contamination have led investigators to acquire fresh stocks of HeLa CCL‐2 cells, but results with the HeLa CCL‐2 cells do not always reproduce results with HeLa cells that have long history in the laboratory. When used for TCID 50 assays of Coxsackievirus B3/28 (CVB3/28), HeLa CCL‐2 cells returned titers for CVB3/28 that were more than ten‐fold lower than titers obtained using laboratory HeLa cells. The viral cytopathic effect was less distinct in the HeLa CCL‐2 cultures, suggestive of a mixed population of cells with varied susceptibility to viral cytopathic effect. Analysis of short tandem repeat markers confirmed the identities of the cell lines as HeLa. Subpopulations in the HeLa CCL‐2 culture, separated easily based on the speed with which they were released by trypsin‐EDTA, differed in their susceptibilities to CVB3/28 cytopathic effect, and in their expression of the Coxsackievirus and adenovirus receptor (CAR). The distinctions between Lab HeLa and HeLa CCL‐2 cells were less obvious when infected with CVB3/RD, a strain selected for growth in RD cells. Results that differ among laboratories may be due to the use of HeLa cell strains with different histories, and experiments using HeLa CCL‐2 available from the American Type Culture Collection are probably incapable of reproducing many of the published studies of Coxsackievirus that have used HeLa cells with laboratory‐dependent histories. J. Med. Virol. 85:677–683, 2013. © 2013 Wiley Periodicals, Inc.