Prevention, Diagnosis and Eradication of Mycoplasma Contamination in Cell Culture

Xuefeng Huang; Minghang Yu; Bingbing Wang; Yanlong Zhang; Junjing Xue; Yu Fu; Xi Wang

doi:10.14440/jbm.2023.407

Prevention, Diagnosis and Eradication of Mycoplasma Contamination in Cell Culture

J Biol Methods. 2023 Nov 1:10:e99010005. doi: 10.14440/jbm.2023.407. eCollection 2023.

Authors

Xuefeng Huang^{1

2

3

4}, Minghang Yu^{1

2

3

4}, Bingbing Wang^{1

2

3

4}, Yanlong Zhang⁵, Junjing Xue^{6

7}, Yu Fu⁶, Xi Wang^{1

2

3

4}

Affiliations

¹ Beijing Key Laboratory of Emerging Infectious Diseases, Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, P.R. China.
² Beijing Institute of Infectious Diseases, Beijing, 100015, P.R. China.
³ National Center for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, P.R. China.
⁴ National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing,100015, P.R. China.
⁵ Department of Urology, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038, P.R. China.
⁶ State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100080, P.R. China.
⁷ Shandong First Medical University & Shandong Academy of Medical Sciences, Shandong, 250000, P.R. China.

Abstract

Mycoplasmas, which are the smallest and simplest prokaryotes, lack a cell wall but possess the ability to undergo self-replication. Mycoplasma contamination is a common problem for laboratories engaging in cell culture. Due to their small size, Mycoplasmas can easily permeate filters designed to prevent bacterial and fungal contamination in cell culture. Although Mycoplasma contamination usually does not result in cell death, it can significantly affect cell proliferation, metabolism, and cause chromosomal aberrations. Therefore, it is crucial to detect and eliminate Mycoplasma contamination in cell culture. This step-by-step protocol presents a comprehensive approach to prevent Mycoplasma contamination in cell culture, as well as to detect and eradicate Mycoplasma to ensure accurate experimental and sequencing results.

Keywords: Mycoplasmas; cell culture; contamination; diagnostic; eradication.