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Salvaging contaminated Matrigel


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5 replies to this topic

#1 kitsonliew

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Posted 10 May 2011 - 06:04 PM

Hi everyone,

I am suspecting some of my small Matrigel aliquots are contaminated. I observed the Matrigel-coated filter under the microscope and none of my cells invaded through the membrane. I can only see bacteria-like stuffs which makes me worry a lot.

If my small Matrigel aliquots are really contaminated, is there any way to salvage it? Is it possible to add serum-free medium supplemented with antibiotic while diluting it to working concentration? Please help as I do not really want to discard them straight away. I would greatly appreciate if there are good tips and ideas other than discarding them.

Thank you very much and warmest regards,

Kitson

#2 bob1

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Posted 10 May 2011 - 07:06 PM

The question you should ask yourself is: Would I use this knowing that bacteria have been growing in it and potentially changing the components (by metabolism etc.)?

#3 kitsonliew

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Posted 10 May 2011 - 07:36 PM

Thanks Bob1 for the reply.. I am still suspecting and my matrigel aliquots are still in - 20 C. So, though it may sound honestly stupid, I am keeping my fingers crossed hoping that extreme low temperature can suppress any microbial activities..

#4 bob1

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Posted 11 May 2011 - 05:02 PM

But what will happen when you go to use the matrigel? If there are bugs in there - throw it out, it's not worth getting a result and then worrying that it might be wrong because of potential contamination.

#5 kitsonliew

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Posted 11 May 2011 - 06:39 PM

Hi Bob1,

Yesterday I tested one of the small Matrigel aliquots for contamination. I diluted the Matrigel with serum-free medium and cultured it overnight. This morning when i took from the incubator to observe, there was no sign of contaminants =). But to be on safe side, I will leave it for another day.

Bob1, have you ever had an experience whereby your Matrigel coat formed a "basin" when it is dry? The gel concentrates and mainly dries on the periphery side of the membrane rather than the centre. As a result, the coating of the membrane was uneven and the cells tend to invade across the membrane centre. Do you have any recommendation or ideas to solve this problem? Thank you very much

Kitson

#6 bob1

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Posted 12 May 2011 - 08:35 PM

Sorry, never used matrigel. I was just applying general lab principles to your problem.




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