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Best practice use of antibiotics


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#1 oliviaw

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Posted 09 August 2010 - 05:32 PM

Hi Guys,
I'm interested in how and why you use antibiotics in cell culture. I have routinely used antibiotics for a while now (twas how I was taught).
But I have learnt the error of my ways and will change my cultures to antibiotic free.
BUT ..
I am wondering if there are times that you do use antibiotics? For example, when you get new cells (from ATCC) and need to get frozen stocks. Or when you bring up cells from frozen stocks.
I would never try and 'clean up' cultures - they get binned instead. Wouldn't trust my results.
Thanks,
Olivia

#2 jessc

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Posted 09 August 2010 - 10:48 PM

I wouldn't generally use antibiotics to bring cells up from thaw, or if you receive new cells from ATCC, they should have strict QC requirements that verify the sterility of any cells they supply. The only time my lab uses antibiotics are if they are primary cell lines or if the cells are really important and don't have back up stocks.

#3 rhombus

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Posted 10 August 2010 - 01:48 AM

Hi Guys,
I'm interested in how and why you use antibiotics in cell culture. I have routinely used antibiotics for a while now (twas how I was taught).
But I have learnt the error of my ways and will change my cultures to antibiotic free.
BUT ..
I am wondering if there are times that you do use antibiotics? For example, when you get new cells (from ATCC) and need to get frozen stocks. Or when you bring up cells from frozen stocks.
I would never try and 'clean up' cultures - they get binned instead. Wouldn't trust my results.
Thanks,
Olivia



Dear Olivia,

Cells in culture should always be antibiotic free. The only exception is when you transfect and use a selection antibiotic. Primary cells are the most difficult cells to grow in general terms. They normally come from an area which is not sterile....for example an abattoir. They are not immortallised so will have a limited life span. You should again never use antibiotics/antimycotics in primary culture...they are very dirty compounds.
One example from years ago:- isloating primary endothelial cells from pig aorta. In the winter good numbers of clean flask's. However in the summer the ambient temperatures rise, causing more contamination. We used fungizone to keep the contamination down. This was successful....BUT the physiological substance released from the cells was NOT present under these conditions. The fungizone had INHIBITED the cellular response we were looking for (unpublished).

In my experience, always CHUCK cells that are contaminated. You can never successfully clean cells up. Also the cells CHANGE in the presence of contamination... and will not revert back. This is particulary imortant when cells are contaminated with MYCOPLASMA.


Hope this helps

Kindest regards

Uncle Rhombus




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