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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

-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

-bob1-

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

-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.

-AussieUSA-