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Position: Discussion forums > Bioscience biotechniques discussion forums > Cell biology and histology methods forum
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EGF culture supplement question??
Posted by: beaker (IP Hidden, Unregistered user, )
Date: April 24, 2005 03:34PM
recently, I culture a epithelial cell, and it's culture media need to be supplemented by EGF. But I want to know how should I add this EGF? whether I add it in media before or after filtering media?I usually store the meida at 4 oC,is it all right that the media supplemented by EGF and serum are stored at 4 oC?
Re: EGF culture supplement question??
Posted by: huangz123 (IP Hidden, Junior member, 19)
Date: April 24, 2005 04:10PM
It's better to add EGF or any other growth factors immediately before use. Aliquote your growth factors and store in -20c per manufacturer's instruction. Calculate amount of medium you may need and add growth factors just before use. This way you will avoid variations of growth factor activity.
Re: EGF culture supplement question??
Posted by: 5'-ATCG (IP Hidden, Unregistered user, )
Date: April 26, 2005 05:20PM
Depend on how soon you consume your medium. If you only use 100-200 mL of medium per week then huangz123's method is prefer. If you need large quantity of medium (>500 mL per week) or need to change medium on more than 20 dishes, I suggest you to add EGF to your medium before filtration.
The medium with serum and EGF supplement can be stored at 4 degree for upto 2 weeks no problem in our lab.
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