how to dissociate cells for cell migration assay
Posted 31 August 2010 - 08:50 AM
I am planning to perform cell migration assays (MCF7) cells. My question is how to dissociate the cells before placing them in the upper chamber for migration? My problem is that if I use trypsin, it needs to be inactivated, but in my assay I need to work with cells that are starved (no FBS)
Posted 01 September 2010 - 01:06 AM
Posted 01 September 2010 - 05:24 AM
Posted 11 September 2010 - 12:35 AM
trypsin can cleave off some cell surface receptors , and u should check if your's does too..
if not, i guess it is fine..
i usually use the citric saline method to harvest my cells,
as the receptor im studying on gets cleaved by trypin..
Citric saline (from Lesley Barber, Melbourne, Australia)
(a) Less harsh than EDTA or trypsin, good viability.
( Recipe for 10X stock solution of citric saline
(i) 1.35 M potassium chloride (KCl, MW=74.55, 10.06 g/100 ml)
(ii) 0.15 M sodium citrate (C6H5Na3O7•2H2O, MW=294.1, 4.41 g/100 ml)
(iii) Autoclave and store at 4 C
(iv) Dilute 1:10 with sterile distilled water before use
(i) Remove all media, flood flask bottom with 1X citric saline
(pre-warmed to 37 C)
(ii) Incubate at 37 C for 4 minutes
(iii) Tap flask bottom to gently detach all cells