Skiet
Posts: 1
Score: 0
Joined: 13/7/2005
Status: offline
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RE: HEK293 Cell Line (in reply to cfpavel)
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Hi,
I regularly use 293's, but the problem you describe i encounter more with 293T's.
With 293's they go really bad when the passage number gets too high. I don't know how often you split them, but here the following procedure works fine...
I split them 2x per week usually a monday and thursday or friday. I always plate out 20% of a full flask in a new flask. I don't trypsinize, instead remove medium from the cells, and "knock" them loose "dry" then respuspend in new medium and plate out in a new flask/dish.
It is possible that you are splitting the cells too heavily, then getting colonies developing and having to leave the cells too long before splitting them again, whilst those in the middle are no longer growing. Once everything starts to die they come loose in "sheets".
Hope that helps a bit, let me know how it goes,
greetings,
Skietje
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