Cell cultures are utilized for various purposes including studies on cell proliferation and the production of biologically active substances. They play a vital role in application development in research laboratories, regenerative medicine, and other biotechnological production platforms. Good cell culture conditions are ideal to achieve reproducible results amid rising concerns about mycoplasma contamination.
Download this infographic now to learn more about reducing the risk of mycoplasma contamination through selecting easy-to-clean pipettes, sterility-promoting consumables, and regular testing.
Cell cultures are utilized for various purposes including studies on cell proliferation
and the production of biologically active substances. They play a vital role in
application development in research laboratories, regenerative medicine, and other
biotechnological production platforms. Good cell culture conditions are ideal to
achieve reproducible results amid rising concerns about mycoplasma contamination.
Detection and Prevention
of Mycoplasma
Contamination in
Cell Cultures
Reduce the risk of mycoplasma contamination and spread in your cell culture
lab with good cell culture practices, selecting easy-to-clean pipettes, sterility-
promoting consumables, and regular testing.
Mycoplasma contamination can
extensively a?ect cell physiology
and metabolism and is prevalent
in cell culture labs. This is because
mycoplasma spread e?ciently even in
routine subculturing. Mycoplasma are
a genus of bacteria that lack a cell wall
and can take on di?erent shapes, from
round to elongated. They are resistant
to antibiotics and due to their small size
(0.2-0.8 μm), can penetrate sterilizing-
grade ?lters, making them very di?cult
to eliminate and spot within cell cultures.
Researchers have identi?ed mycoplasma contamination in 11% of 10,000 cell
lines tested (Olarerin-George et al. 2015) with live mycoplasma also detected
outside of the flask, on a hemocytometer, on the pipette, and outside of the
pipette discard pan after working with infected cell cultures in a laminar flow
hood. Live mycoplasma were also recovered from the surface of the laminar
flow up to six days after the initial detection.
What can mycoplasma do to
cell cultures?
Mycoplasma can inhibit protein biosynthesis and cell growth and alter RNA and DNA
synthesis. In cell-based therapeutics and in advanced therapy medical products
(ATMPs), mycoplasma contamination can cause immune reactions, chromosomal
aberrations or altered proliferation characteristics. This can make the resultant
therapies unsafe for use in patients (Martins et al. 2014).
Potential routes of mycoplasma contamination include:
(i) Contaminated media, reagents
or instruments. Pipettes are
frequently exposed and must be
fully autoclavable or have dispensing
head parts that can be autoclaved.
(ii) Laboratory personnel (~80.6% of
technicians are carriers)
(iii) Infected cultures obtained from
other labs
Practices to avoid mycoplasma
contamination
1. Implement good cell culture practice
2. Select easy-to-clean and autoclavable pipettes for cell culture work
3. Select sterile ?lter tips with protective wrappings
4. Regularly test all cell lines (especially new cell lines) for contamination
5. Use 0.1 μm sterile ?lters for ?ltering cell culture reagents and media
Solutions for detecting and
preventing mycoplasma
contamination
Sartorius’ Microsart
®
AMP Mycoplasma qPCR Mycoplasma and Microsart
®
ATMP
Mycoplasma detection kits provide for early detection of mycoplasma contamination.
Fast, highly speci?c, sensitive, and compliant with international guidelines, Microsart
®
Mycoplasma qPCR detection kits enable e?ective detection of mycoplasma
contamination in your cell culture samples.
Sartorius’ Microsart
®
AMP
Mycoplasma and ATMP qPCR
Mycoplasma Detection Kits
• High performance EP compliant Mycoplasma
testing with detection limit of less than 10
cfu/mL for all mycoplasma species in the
European and U.S. Pharmacopeia
• Fast-real time PCR kit with DNA extraction to
PCR results taking only a few hours
• TaqMan
®
probes provide speci?city to PCR
detection system with no melting curve
analysis needed
• Flexible samples volumes from 200 µL to 18 mL
• Minimal false-positive results as kit contains
dUTP instead of dTTP with option to degrade
amplicons from prior analyses using uracil-
DNA glycosylase (UNG)
These kits are supplemented by the use of Sartorius’ single-channel Tacta
®
pipettes
that cover a volume range of 0.1µL – 10,000 µL, with multichannel pipettes available
from 0.5 µL to 300 L to 300 µL volume, and Minisart
®
NML 0.1 μm-5 μm pore size
syringe ?lters for optimal clari?cation and sterilization of liquids, robust removal of
bacteria and particles, with minimal environmental impact.
Proper awareness of mycoplasma sources and the use of aseptic cell culture
techniques supplemented by reliable detection methods for mycoplasma
contamination can help prevent and reduce the risk of mycoplasma
contamination in cell cultures. Sartorius’ Microsart
®
AMP Mycoplasma qPCR
Mycoplasma detection kits alongside their single-channel Tacta
®
pipettes and
Minisart
®
NML syringe ?lters provide an accurate, precise, fast, and convenient
means for mycoplasma contamination detection and prevention to support
your lab’s cell culture needs.
To learn more, visit www.sartorius.com
Sartolab
®
Vacuum Filters
Filtration of cell culture media, especially
media prepared from powder, represents
a cornerstone of a comprehensive
prevention strategy. Sartolab
®
RF
vacuum ?lters with a 0.1-μm PES
membrane can e?ectively remove
Mycoplasma from cell culture media.