I am growing HEK293 cells (adhered) in serum free media. Day 1 afer passaging, cells grow in monolayer, but second day onwards cells start forming clusters on monolayer.
I tried to make fine suspension using pipette for a long time but this is causing mechanical shear to cells. Any suggestions please?
Hello Sonika
From your question what I have understood is that are you trying to adapt adherent HEK293 into suspension culture? Because you mentioned that you are using serum free medium.
You will have to culture adherent HEK293 with decreasing concentration of fetal bovine serum over time starting with 10% FBS, followed by 5%FBS and so on till you reach serum free condition. This will take a while for HEK293 to get adapted to serum free medium.
HEK293 in suspension culture also form clumps. To overcome this problem anti-clumping agent is used.
Regards.
@Malcolm Nobre,
Thank you for your reply!
I am not trying to adapt HEK293 in suspension. I am growing them in adherent only in Free Style 293 media (serum free). But cells are forming round clusters over monolayer ( spindle shape). Any suggestions?
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