I want to culture my cancerous cell line in serum free media to detect cancer stem cells by flow cytometry. How many days before doing flowcytometry would be suitable to culture them to be able to detect cell surface markers?
Two days prior to flowcytometry you have to seed your cell line, also depends on the time period of growth of the cell line which is in how many days your cell line forms a mono layer before you can trypsinze and collect them and then do the processing for flowcytometry.
Hi, Tarlan,
this is largely dependent from the cell types you have used, from the "history" and growth-ratio of your cell lines.
For rapid growing cells 1.5-2 days will be enough.
There is a good protocol for stem cell detection with Hoechst, Plesae, follow it!
Good luck!
http://antibody-antibodies.com/product1947852-TNCbio-XerumFree%E2%84%A2.html
http://gentaur-worldwide.com/images/datasheets/XerumFree.pdf
did you try xerumfree??
Defreeze your culture in growth medium for 3 days check cell viability by Trypan blue ; 48H before the assay change the growth medium to serum-free medium. Good luck!
Hello Tarlan,
it really depends on the cell line you re using. But notice that if you leave your culture for a long periode, and even in a serum free media your cell line will start to differentiate and you might loose some markers. We already experienced this in our studies you can check our last article. It might give you some ideas.
http://www.biomedcentral.com/content/pdf/1746-6148-9-224.pdf
good luck!!
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