How to deal with mycoplasma contaminant in cell cultures? The growth of my cells was obviously hampered.
Dear Mei
Mycoplasma detection can be perfomed by simple PCR assay
(you can find a protocool of it on my blog: http://proteocool.blogspot.com
at PAGE 5
P roteoCool n°18 Simple PCR test to detect Mycoplasma contaminations
positive sample need to be treated with specific antibiotics. Several antibiotics are avialable (eg, B-M ciclyns , cyprofloxacins and others)
Article Treatment of Mycoplasma Contamination in Cell Cultures with Plasmocin
generally in our lab we perform two cicle of BM ciclyn treatment and at the end we re-test the presence of mycoplasma with PCR methods and in the most cases the problem is solved.
You can reduce the risk of mycoplasma contamination using 0,1uM filter for media preparation insted 0,22uM but do not eliminate the risk.
i hope that this informations may be usefull to you
ciao
MAnuele
Dear Lan Mei,
To go further you can also give a look at this article. It compares efficiency of different mycoplasma removal reagents among time. Mycoplasma detection was performed using nuclear staining by Hoechst 33342 and DNA gel electrophoresis.
Hope this helps !
Nicolas
Article Evaluation of mycoplasma removal reagents using qPCR-based q...
Recovering cells from mycoplasma is critical even though its recovered partially its not good to experimentation since physiology has changed.
Through the all cultures, fumigate the lab. Start freshly culturing the cells in media containing antimycotic.
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