Subculturing without trypsin - (May/27/2008 )

Hi all,

The cell lines I'm working with (MCF7, HEK293) detach easily by pipetting a few times without trypsin. Therefore, I am omitting trypsin while subculturing. Is there a disadvantage to this? I have not encountered any specific problems but I'm new to cell culture so I want to know if this is good practice.

Thanks,
Solanacea

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1. Over-enthusiastic pipetting may damage and subsequently kill many cells.
2. The cells may remain as cell-clusters and not individual cells which might be desirable for some experiments -uniformly spread, fast confluence.
3. Otherwise perfectly fine. We routinely subculture 293t like that, which is one of the easiest to detach.

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since trypsination is a harsh method to detach cells, pipetting is to prefer; you may support pipetting by adding EDTA in PBS

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