cell detachment - help (Dec/15/2005 )

Helloo
when i try to detach my cell lines from the culutre flask using trypsine but all the cells round off and form a mass, could anyone help me to over come this problem
Best regards
cell line

Hi

Do you mean that they form clusters of cells? or one big mass? If it is just clusters, try trypsinising for a slightly shorter time and gently pipetting up and down a few times to break any clusters (how hard you can do this depends on the cell lines, hela and A549 are really tough, but embryonic fibroblasts are more fragile).

Bob

Dear Bob
thanks for the reply.. it actually forms a big mass rather than clustering. Looking forward ur suggestions
Best regards
cell line

The presence of EDTA (~0.5 mM) helps keep clumping down, either with the trypsin (i.e. trypsin:EDTA or versene) or by it's self.