EDTA toxicity when detaching adherent cells SUMMARY

Guillermo Palao gpbnas at yahoo.es
Tue Aug 30 04:58:02 EST 2005


Hello all,


I received many answers on the best way to detach adherent cells (primary human
fibroblasts) with EDTA, so I thought it might be interesting to post a summary of the
advices, which have been tremendously helpful:

- Best EDTA temperature was 37ºC, at 4ºC cells barely detached at all. Washing with PBS
before EDTA greatly shortened time needed for detachment of cells.

- Adding proteins, in this case serum (10% FCS) to EDTA (0.6 mM) markedly improved
viability without increasing time to detachment.

- Interestingly, even T.Blue positive cells after EDTA treatment where successfully
subcultured, so a transient increase in cell permeability did not mean irreversible cell
death. 


Thank you all again,



Guillermo Palao, MD
Laboratorio de Reumatología
Centro de Investigación
Hospital 12 de Octubre
Madrid
Spain
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