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tissue collection and transport medium


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#1 Johannes

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Posted 15 September 2013 - 11:06 PM

Hi there,

 

I am planning to collect bovine tissue samples at an abattoir and was wondering what medium would be best to collect the samples in. I then want to mince and digest these samples and try to establish primary cell cultures. I did some reading and found all kinds of combinations of media, buffer, antibiotics and fungicides being used, but nothing specific so far. I remember in my old lab they used Hanks medium to collect human samples from the pathology lab. But as it won't be that sterile in the abattoir I was thinking of supplementing the medium with pen/strep.

 

Does anyone have any advice on antibiotic concentrations or media/buffer I should use in my transport medium, or even better a protocol for tissue collection and further processing in cell culture?

 

Thank you very much in advance,

 

Johannes



#2 bob1

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Posted 16 September 2013 - 12:21 PM

I would suggest first washing the specimens in PBS or something similar, then dipping in 70% EtOH briefly (less than 1 min), the rinse again in PBS a few times.  This should kill most of the surface bacteria.  The specimens can then be transported in whatever medium you prefer, PBS or Ringer's buffered saline would be good options. 

 

The primary culture, it will pay to assess the use of antibiotics frequently, you may well find that they are unnecessary, especially the anti-fungals.



#3 Johannes

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Posted 16 September 2013 - 05:38 PM

hey bob,

 

I avoid using antibiotics in my cell culture, I just thought it might be a good idea for the transport medium, as I will probably not be the one collecting the specimen. 

Thanks for your  tips. I will advice the person who collects to do it like that, and then assess the samples in the lab for sterility. 



#4 rhombus

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Posted 26 September 2013 - 07:21 AM

Dear Johannes,

 

I used to collect Porcine Aorta from abattoirs in the 1980's to isolate Endothelial cells. What I found was that the fresh aortae were happy to be in a dry 2L beaker for the transportation in a cool box (up tp 3 hrs).What you will find is that if you use a "media" of some sort, you will just transfer contamination from one aorta/tissue to another. However much you wash the aorta/tissue with PBS, once contaminated they will never be clean. Using strong antibiotics/antimycotics may help to reduce contamination BUT these compounds themselves have a massive effect on the cells.

 

Hope this is useful

 

Uncle Rhombus






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