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J Cell Mol Med. 2019 Jan;23(1):227-236. doi: 10.1111/jcmm.13923. Epub 2018 Oct 24.

Rapid detection of low-level HeLa cell contamination in cell culture using nested PCR.

Lin J1,2,3, Chen L1,3, Jiang W1,3, Zhang H1,3, Shi Y1,3, Cai W1,3.

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Institute of Apply Genomics, Fuzhou University, Fuzhou, China.
School of Basic Medical Sciences, Fujian Medical University, Fuzhou, China.
College of Biological Science and Engineering, Fuzhou University, Fuzhou, China.


HeLa cells are a commonly used cell line in many biological research areas. They are not picky for culture medium and proliferate rapidly. HeLa cells are a notorious source of cell cross-contamination and have been found to be able to contaminate a wide range of cell lines in cell culture. In this study, we reported a simple and efficient method for detecting the presence of HeLa cell contamination in cell culture. HPV-18 was used as a biomarker. The cell culture supernatant was used directly as the template for nested PCR without extracting nucleic acid. By PCR amplification of the cell culture supernatant with the designed primers, we were able to detect the presence of HeLa cells in the culture. The sensitivity of this method can reach 1%, which is 10-fold higher than Short tandem repeat sequence (STR) profiling. This simple, rapid, and "noninvasive" quality checking method should find applications in routine cell culture practice.


HeLa; cell cross-contamination; nested PCR

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