Protocol Online logo
Top : New Forum Archives (2009-): : Cell Biology

A549 cells are not attaching to flask - (Jun/24/2012 )

I am new to cell culture , once I thawed the cells and seeded in RPMI ( 10%FBS and 1% p/s) they are not attach to the corning flask . I Don"t know why ??????????? I tried with several times. but it gives same results .I followed extreme aseptic conditions . But i am not sure with incubators contamianated or not . Pls help me


Check if your cells are alive or dead (trypan blue stain and haemocytometer).

How long did you leave them before deciding they were not attaching?
Is there someone in the lab who is also growing the cells and they are fine?
Have you had someone show you how to thaw cells properly?
Are you using the medium that these cells have been grown in before freezing?


according to this,
you seemed to have used the wrong medium. but anyway, sometimes we dont follow exactly the protocol from ATCC. regarding the medium, if a specific one is not available, usually we use DMEM for adherent cells and RPMI for lymphoid or suspension cells. If I were you, i would try DMEM first.