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How to culture stable cell line?(urgent help needed)
Posted by: popogirl (IP Hidden, New member, 9)
Date: April 13, 2006 11:43PM
Hi,
I am very new in culturing stable cell line. Recently I ask a kind of stable cell line from someone. I thraw them and let them grow about 10 days and suddenly realize that I shoud add G418 to it. So I add 300ug/ml G418 to the cell immediately. Could anyone tell me is that OK or it will cause me lose the stable expressed gene? How long it will take for G418 to kill the cells that do not express the gene stably?How to maintain the gene stably expressed in cell? Should I add G418 immediatly when I thraw the cells or wait for several days to let the cell recovery and then add drug? Thank you very much!
Re: How to culture stable cell line?(urgent help needed)
Posted by: mitolab (IP Hidden, Senior member, 89)
Date: April 25, 2006 10:27AM
Since you are just maintaining the stable cell line I think it should be fine to add G418 after few days of thawing.
Re: How to culture stable cell line?(urgent help needed)
Posted by: fatboym (IP Hidden, Junior member, 20)
Date: April 27, 2006 04:14AM
well Mitolab is right but I wonder if the concentration you use is appropriate for your cell line, since cancer cell lines are usually maintained with higher concentrations
Re: How to culture stable cell line?(urgent help needed)
Posted by: dkhurtado (IP Hidden, New member, 5)
Date: June 7, 2006 02:16PM
What kind of cell line are you culturing? Different cell types require different concentrations of G418. My experience ( 15 years in a cell culture lab) has been that Hek293 - 750ug/ml,
CHO - 500ug/ml for example. Its fine to grow the new thaw without antibiotics for at least one week - they need time to recover.
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