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

Jun Lin; Lin Chen; Wenqian Jiang; Huilian Zhang; Yang Shi; Weiwen Cai

doi:10.1111/jcmm.13923

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

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

Authors

Jun Lin^{1

2

3}, Lin Chen^{1

3}, Wenqian Jiang^{1

3}, Huilian Zhang^{1

3}, Yang Shi^{1

3}, Weiwen Cai^{1

3}

Affiliations

¹ Institute of Apply Genomics, Fuzhou University, Fuzhou, China.
² School of Basic Medical Sciences, Fujian Medical University, Fuzhou, China.
³ College of Biological Science and Engineering, Fuzhou University, Fuzhou, China.

Abstract

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.

Keywords: HeLa; cell cross-contamination; nested PCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Culture Techniques / methods*
Cell Line
HeLa Cells*
Human papillomavirus 18 / genetics
Humans
Microsatellite Repeats
Polymerase Chain Reaction / methods*
Reproducibility of Results

Associated data

GENBANK/NC_001357