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Aortic smooth muscle cell contamination

Cell culture Contamination Smooth Muscle Primary culture

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#1 craiganash

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Posted 02 July 2012 - 12:18 AM

I currently culture both aortic and mesenteric primary smooth muscle cells from rat. The Aortic smooth muscle is cultured in DMEM + 10% FCS, 10ug/ml Pen/Strep and 10ug/ml amphoteracin B. However the cells from the aorta have recently been getting infected periodically two days into culture. Interestingly, this does not happen to the mesenteric cells.

I have tried all the usual solutions to remove cell culture contamination, such as cleaning incubators and hoods with 10% microsol and performing the dissection in a sterile manner, with antibiotics in the dissection solutions, however, none of this is helping. The contaminant has been identified as a bacillus type bacteria by a bacteriologist in our Immunology dept. Does anyone have any suggestions to help with this problem?

Many thanks

Edited by craiganash, 02 July 2012 - 12:19 AM.


#2 bob1

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Posted 02 July 2012 - 01:07 PM

The bug could easily be in your stocks of cells at a very low concentration if you are using thawed stocks.

Silly question, but have you thrown out all your currently open culture medium/PBS/trypsin/collagenase/antibiotics/etc? If not, that would be the first place to start.

Make sure you sterilize your dissection equipment by autoclaving and only open in the hood.





Also tagged with one or more of these keywords: Cell culture, Contamination, Smooth Muscle, Primary culture

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