Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Cell Biology and Cell Culture
Register Search Today's Posts Mark Forums Read

Cell Biology and Cell Culture Cell Biology Forum. Cell Culture Forum. Post and ask questions about cell culturing, cell lysis, cell transfection, cell growth, and cell biology.


Mycoplasma contamination?

Mycoplasma contamination? - Cell Biology and Cell Culture

Mycoplasma contamination? - Cell Biology Forum. Cell Culture Forum. Post and ask questions about cell culturing, cell lysis, cell transfection, cell growth, and cell biology.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 09-11-2009, 05:14 AM
Pipette Filler
Points: 51, Level: 1 Points: 51, Level: 1 Points: 51, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2009
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Default Mycoplasma contamination?



Hi everyone,
My PC12 cells frequently got contaminated recently. I usually use 320x invert microscope to look at the cells, and if they are contaminated by conventional bacteria (with cell wall) I can clearly see those "rods". But these days the medium remains clear for 5-6 days and then suddenly turns turbid overnight. I observe the flask under 320x magnification again, but cannot see the usual bacteria. Instead I vaguely see very tiny "-"shaped vibrating things. They are densely present in the culture, while the PC12 cells detach and I suppose they're dead. I suspect if it's mycoplasma contamination? Many websites say they can be hardly detected by conventional microscopy, so I doubt whether what I saw are really mycoplasma... Has anybody actually seen the mycoplasma before?
Thanks in advance for any comments!
Reply With Quote
  #2  
Old 09-11-2009, 09:35 AM
Pipette Filler
Points: 41, Level: 1 Points: 41, Level: 1 Points: 41, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2009
Posts: 3
Thanks: 0
Thanked 2 Times in 2 Posts
Default Re: Mycoplasma contamination?

Hi!

We have had mycoplasma infections in our cell cultures quite often. It keeps coming back, unfortunately... But we have never ever seen it. We detect it using PCR.

Thessi
Reply With Quote
The Following User Says Thank You to Thessi75 For This Useful Post:
admin (09-14-2009)
  #3  
Old 09-13-2009, 06:00 AM
Pipette Filler
Points: 51, Level: 1 Points: 51, Level: 1 Points: 51, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2009
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Mycoplasma contamination?

Hi Thessi, do you observe turbid medium if your cell cultures are contaminated with mycoplasma? And would it be so serious that the cells all die, or they just coexist? Thanks for your response!
Reply With Quote
  #4  
Old 09-14-2009, 10:49 AM
Pipette Filler
Points: 41, Level: 1 Points: 41, Level: 1 Points: 41, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2009
Posts: 3
Thanks: 0
Thanked 2 Times in 2 Posts
Default Re: Mycoplasma contamination?

Hi!

The cells tend to look normal and so does the medium. Cells that have tested positive for mycoplasma seem to coexist well with the infection (we have leukemic cell lines in our lab). We test and treat all our cell cultures for mycoplasma regularly.

From time to time our cells have looked rather unhappy, but we are not sure whether it has been due to the presence of mycoplasma or not. Recently I experienced that several cell cultures just more or less disintegrated (lots of dead cells and very turbid medium), without any apparent infection.

Hope you'll get rid of whatever is growing in your cultures. Soon!

Thessi
Reply With Quote
The Following User Says Thank You to Thessi75 For This Useful Post:
admin (09-14-2009)
  #5  
Old 09-17-2009, 08:56 PM
Pipette Filler
Points: 44, Level: 1 Points: 44, Level: 1 Points: 44, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2009
Posts: 6
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Mycoplasma contamination?

There are some kits and even easy DAPI staining for detecting mycoplasma in your cell cultures.
You can search on main commercial life science providers what product is the best suitable for your lab.
Reply With Quote
  #6  
Old 09-17-2009, 08:56 PM
Pipette Filler
Points: 44, Level: 1 Points: 44, Level: 1 Points: 44, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2009
Posts: 6
Thanks: 0
Thanked 0 Times in 0 Posts
Wink Re: Mycoplasma contamination?

There are some kits and even easy DAPI staining for detecting mycoplasma in your cell cultures.
You can search on main commercial life science providers what product is the best suitable for your lab.
Reply With Quote
  #7  
Old 09-18-2009, 09:10 AM
Pipette Filler
Points: 2,304, Level: 31 Points: 2,304, Level: 31 Points: 2,304, Level: 31
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jun 2009
Posts: 9
Thanks: 0
Thanked 1 Time in 1 Post
Default Re: Mycoplasma contamination?

I've had a similar issue with the cultures once. I doubt it is micoplasma, it is usually barely noticable and often coexists with cultures. Often the only way you know something is wron, is when your experiment control down't work as supposed to. I would say scrap the cultures, clean/autoclave the incubator, get new media, new sera, etc, and get new cells from ATCC. Do you use antibodies in your media?
Good Luck!
Reply With Quote
The Following User Says Thank You to garlic For This Useful Post:
admin (09-18-2009)
  #8  
Old 09-18-2009, 09:11 AM
Pipette Filler
Points: 2,304, Level: 31 Points: 2,304, Level: 31 Points: 2,304, Level: 31
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jun 2009
Posts: 9
Thanks: 0
Thanked 1 Time in 1 Post
Default Re: Mycoplasma contamination?

I've had a similar issue with the cultures once. I doubt it is micoplasma, it is usually barely noticable and often coexists with cultures. Often the only way you know something is wron, is when your experiment control down't work as supposed to. I would say scrap the cultures, clean/autoclave the incubator, get new media, new sera, etc, and get new cells from ATCC. Do you use antibodies in your media?
Good Luck!
Reply With Quote
  #9  
Old 09-20-2009, 01:34 PM
Pipette Filler
Points: 51, Level: 1 Points: 51, Level: 1 Points: 51, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2009
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Mycoplasma contamination?

Thanks everybody, now I guess it's normal bacteria since the medium gets really turbid and cells all die... I did use 0.5% PSN but maybe that's not enough? Anyway I've cleaned the incubator and made new medium, now waiting for new cells from the postgrads in my lab.
Reply With Quote
  #10  
Old 01-21-2010, 08:31 PM
Pipette Filler
Points: 185, Level: 3 Points: 185, Level: 3 Points: 185, Level: 3
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jan 2010
Posts: 5
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Mycoplasma contamination?

I've used the mdbioproducts PCR mycoplasma detection kit with success.

This company has also had good technical support when I needed help or had questions.

Just google their name to find them online.
Reply With Quote
Reply

Tags
contamination , mycoplasma


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Detection of viral contamination? tigerGS08 Cell Biology and Cell Culture 4 02-22-2012 07:51 PM
Contamination of 7H11 agar plates martin rao Microbiology Forum 4 10-31-2010 08:39 PM
Mycoplasma contamination? wangleicau Cell Biology and Cell Culture 3 11-20-2009 01:45 AM
Detecting Mycoplasma Cell Culture Contamination dnamolecule Cell Biology and Cell Culture 9 04-12-2008 07:40 AM
Methods Digest, Vol 21, Issue 25 Jayanta Tarafdar Protocols and Methods Forum 0 02-23-2007 05:46 PM


All times are GMT. The time now is 07:21 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.16752 seconds with 16 queries