P REVENTING C ELL C ULTURE

C ONTAMINATION
NC State Environmental Health & Safety Center / Campus Box 8007 / 2620 Wolf Village Way, Raleigh, NC 27695 / www.ncsu.edu/ehs

Introduction
Cell culture is increasingly prevalent and
important in academic research, and along
with improved proficiency of cell culture
comes a better understanding of the risks and
consequences of cell culture contamination.
Testing methods and preventive tools are
continuously improving, and an awareness of
the effects of contamination requires
researchers to be conscientious and vigilant.
Undetected contamination can lead to widespread downstream effects. Therefore, it is
important to understand how contamination
can occur and what methods are available to
prevent it.

Even a tiny amount of liquid can

cause contamination - one drop of
media left on a bottle thread can
have a detrimental effect on your
resulting data.
detect. Mycoplasmas can be the most difficult
to detect and are resistant to antibiotics. Once
detected, contamination can be contained and
eliminated, but continuous testing is required.

Biological contamination includes bacteria,

fungi, Mycoplasmas, viruses, and crosscontamination from other cells. Bacteria and
fungi are the most common contaminants,
because they are ubiquitous and able to
colonize quickly. They are also the easiest to

Common Sources of Contamination

in Labs
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LABORATORY PRACTICES
Aerosolization of particles
Sneezing, coughing, or stirring up
dust while moving about the lab
Pipetting, vortexing, centrifuging
without bio-containment vessels
Working with multiple cell lines
concurrently
Continuous culturing of working banks
Overcrowding of items in the autoclave
Mixing solutions and supplies

LABORATORY EQUIPMENT
Reservoirs for microbes and fungi
Improperly cleaned incubators
Water baths, refrigerators,
microscopes, cold storage rooms
Use of feeder cells
Culture media, bovine sera, reagents, and
plasticware
Use of non-sterile supplies, media, or
solutions
Materials not certified for cell culture use
Contaminated clothing

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Preventing Cell Line Contamination

A contaminated cell
line can jeopardize
resulting data, create
health hazards to lab
workers, damage
researcher credibility,
and waste resources.

An example scenario of potential cell contamination.

What Can You Do To Prevent Contamination?

Effective training and supervision by principal investigators is critical. Regular use of antibiotics
is discouraged, because it can mask habitual mistakes or poor techniques. The best prevention
against cell contamination is ensuring good laboratory practices among all personnel.
Documentation of all cell culture work
Each passage, manipulation, and
general cell appearance
Ensure proficiency of lab personnel

Label flasks legibly

Include name of cell line, passage
number, date of transfer
Use barcoded flasks when possible

Proper aseptic techniques

Allow >5 minutes for air to be filtered

Aliquot stock solutions/reagents

between working with each cell line

Use one reservoir of medium per cell line

Quarantine dirty cell lines from clean

Work with one cell line at a time

cell lines

Work in a biological safety cabinet

Clean laboratory regularly

Manageable work load

Periodically review and approve lab

Use seed stock

notebooks

Further Reading:
Cases of Mistaken Identity, Science, Vol315, Feb 16, 2007. http://www.sciencemag.org/content/
315/5814/928.full
Eradication of cross-contaminated cell lines: A call for action. R. M. Nardone, Cell Biol Toxicol.,
2007 Nov;23(6):367-72. Epub 2007 May 24. http://www.ncbi.nlm.nih.gov/pubmed/17522957
Cell Culture Contamination, Lab Manager Magazine, Vol6, No4, May2011. http://www.labmanager.com/laboratorytechnology/2011/05/cell-culture-contamination?fw1pk=2#.VAYrImOM2f4
NC State Environmental Health & Safety Center
Campus Box 8007 / 2620 Wolf Village Way, Raleigh, NC 27695 / www.ncsu.edu/ehs