Cell culture-biotechnology--->HELP!

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pinkypig
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Cell culture-biotechnology--->HELP!

Post by pinkypig » Tue Jan 13, 2009 11:08 pm

Can some one explain or suggest a good site which will explain what EDTA is? In a lab experiment we used trypsin:EDTA to dislodge cells from the substratum but i dont know why EDTA was used aswel as trypsin?
Can someone suggest any ideas?

Thanks

blcr11
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Post by blcr11 » Wed Jan 14, 2009 7:14 am

Just google "trypsinization" and you will get most of the information you want to know. EDTA is a standard chelator of (forms a soluble complex with) calcium and magnesium ions, both of which inhibit the enzyme trypsin. Cells themselves contain these metal ions and calcium and magnesium salts are usually added to culture media to the tune of maybe 1-5 mM. By adding EDTA to the media you reduce or remove these inhibitors. Secondarily, there are multiple membrane-associated proteases that are activated by metal ions. These enzymes might be stimulated when trypsin starts to digest away at the cell surface proteins. Trypsin is not "harmless" to cells. You want the enzyme to act only long enough to disrupt the cell-substrate attachments and then you want to get rid of the trypsin/EDTA by dilution and re-plating just as soon as you can. Longer digestions will eventually kill the cells.

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Sepals
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Re: Cell culture-biotechnology--->HELP!

Post by Sepals » Fri Jan 23, 2009 2:58 pm

There's a definition for it on this site:

http://www.biology-online.org/dictionary/Edta

Here's why it's important:

http://www.molecularstation.com/forum/c ... ypsin.html
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