Hello
Have any one tried using the EDTA solution instead of trypsin in detach the adherent cell, in flow cytometry assay, particularly in High throughput analysis
Regards
Hi Asma,
It all depends on how sticky are your cells. Typically for HEK cells it works just fine - we use Versene (EDTA based solution) 10 min at 37oC and then gentle pipeting and they all detach well. But if your cell line is really sticky then it becomes much more difficult to detach cells without damaging them.
we have used Accutase from Sigma. It is EDTA-based (0.5mM) but also has proprietary enzymes of undeclared origin!! so it may not be useful for some applications. For our purposes, it removed sticky cells effectively and rapidly without apparently affecting cell surface protein expression and function.
http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Datasheet/6/a6964dat.pdf
I've found that this has led to huge amount of cell death in lines that I've tried this in. As mentioned already, more strongly adherent lines respond poorly to this method
Thanks all for kind reply
Actually I would like to minimize the washing and centrefugation steps, which are affecting my cell and reading as well
I am working with MCF-7 and LnCaP cell line
What about Accutase solution, is there any one work with these tow cell lines and how it is work
I am Working on 96 well plate , i want to minimize these steps as possible
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