Hey,
I'm planning to use BrdU to label cancer cells in vitro, and inject the cell in vivo, in order to track the cell. For this, I want to know whether BrdU affects the biology of cancer cell or not..(especially metastasis)..I have done some searching on this, but I couldn't find any...
The only thing I found is a study which used cell tracker for this,[Mol Cancer. 2015 Dec;14(1):282] , These guys have done exactly what I am trying to do.
;however, as I know, fluorescent of cell tracker only lasts few weeks,,
That's why I am concerning BrdU
Q1. Would BrdU affect biological behaviour of cancer cells?
Q2. Does anybody know any study using BrdU-treated cancer cells for metastasis experiment?
Q3. Cell tracker vs. BrdU, for this experiment which one would you recommend ?
Thank you!
Yes. Because BrdU intercalates into DNA it can lead to DNA damage and consequently to changes in cellular behaviour. For tracking cells long-term, your best bet is to use a fluorescent dye such as CFSE or SNARF that bind protein. With each cell division after labelling, the amount of stain remaining is halved allowing you to estimate the rate of proliferation over time. Labelling with either of the above reagents is totally straightforward, just be sure to wash away any unincorporated dye before proceeding with your experiments. Good luck!
Yes. Because BrdU intercalates into DNA it can lead to DNA damage and consequently to changes in cellular behaviour. For tracking cells long-term, your best bet is to use a fluorescent dye such as CFSE or SNARF that bind protein. With each cell division after labelling, the amount of stain remaining is halved allowing you to estimate the rate of proliferation over time. Labelling with either of the above reagents is totally straightforward, just be sure to wash away any unincorporated dye before proceeding with your experiments. Good luck!
Yes
Please read these articles
Because BrdU can replace thymidine during DNA replication, it can cause mutations, and its use is therefore potentially a health hazard.[3] However, because it is neither radioactive nor myelotoxic at labeling concentrations, it is widely preferred for in vivo studies of cancer cell proliferation.[6][7] However, at radiosensitizing concentrations, BrdU becomes myelosuppressive thus limiting its use for radiosensitizing.[2]
Konishi, Teruaki; Takeyasu, Akihiro; Natsume, Toshiyuki; Furusawa, Yoshiya; Hieda, Kotaro (2011). "Visualization of Heavy Ion Tracks by Labeling 3'-OH Termini of Induced DNA Strand Breaks". Journal of Radiation Research 52 (4): 433–40. doi:10.1269/jrr.10097. PMID 21785232.
Jump up ^ Kee, N; S Sivalingam, R Boonstra, J.M Wojtowicz (March 2002). "The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis". Journal of Neuroscience Methods 115 (1): 97–105. doi:10.1016/S0165-0270(02)00007-9. PMID 11897369.
Regards
why you don't stably express GFP or mCherry in your cells? metastase need weeks/ months to occur and you will lost your alive markers with time...
I think BrdU and thymidine incorporation assays are mainly suitable for cancer cell proliferation. I think Valentine is right to say use GFP tagged stably protein as live marker for your work.
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