MembersOnly.DetachCells History

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March 29, 2009, at 03:28 PM by 130.126.231.65 -
Changed line 1 from:
'''How to detach the cells from flask while retaining cell surface cadherins'''
to:
'''How to detach MDCK cells from flask while retaining cell surface cadherins'''
March 22, 2009, at 12:06 AM by 98.212.146.133 -
Changed lines 2-4 from:
---------From Johan
Trypsinize MDCK in 0.05% trypsin +EDTA (2mM, standard from GIBCO), wash them first in PBS+4mM EDTA. Here lose all cadherins from the membrane.

Then let the cells recover from trypsinization in suspension (in DMEM + 10% FCS, about 10 Mln cells/ml) for 1 hr, after that wash once in DMEM + 0.5% FCS and resuspend in DMEM + 0.5% at the right concentration for use.
to:
                                                     ---------From Johan
* Wash them first in PBS +4mM EDTA.
* Trypsinize MDCK in 0.05% trypsin +EDTA (2mM, standard from GIBCO), Here lose all cadherins from the membrane.
* Then let the cells recover from trypsinization in suspension (in DMEM + 10% FCS, about 10 Mln cells/ml) for 1 hr
* Wash
once in DMEM + 0.5% FCS and resuspend in DMEM + 0.5% at the right concentration for use.
March 22, 2009, at 12:05 AM by 98.212.146.133 -
Added lines 1-4:
'''How to detach the cells from flask while retaining cell surface cadherins'''
---------From Johan
Trypsinize MDCK in 0.05% trypsin +EDTA (2mM, standard from GIBCO), wash them first in PBS+4mM EDTA. Here lose all cadherins from the membrane.
Then let the cells recover from trypsinization in suspension (in DMEM + 10% FCS, about 10 Mln cells/ml) for 1 hr, after that wash once in DMEM + 0.5% FCS and resuspend in DMEM + 0.5% at the right concentration for use.

Page last modified on March 29, 2009, at 03:28 PM