title: Flow cytometric method for estimation of 5-bromo-2´-deoxyuridine content in rat serum.
abstract: Labelling of DNA in replicating cells using 5-bromo-2´-deoxyuridine (BrdU) is widely used, however the rapid clearance and metabolisation of BrdU in the living organism is a critical issue. Although the pharmacokinetic of BrdU in experimental animals is empirically approximated, the exact time-curve remains unknown. Here we present novel method for estimation of the BrdU content in the blood serum. The application is based on the in vitro cocultivation of tumour cells with the examined serum and the subsequent quantification of the incorporated BrdU in the DNA using flow cytometry analysis. Our results demonstrate that this approach can quantify the BrdU concentration in serum at 1 micromol.dm(-3) and might represent an attractive alternative to conventional chromatographic analysis. The employment of tumour cells as "detectors" of the BrdU content in serum provides an advantage over high pressure liquid chromatography (HPLC), as this approach allows us to approximate not only the concentration of BrdU, but also to determine, whether BrdU is present in the blood serum in effective concentration to reliable label all cells undergoing the S-phase of the cell cycle. The presented application might be a helpful tool for studies on pharmacokinetics of BrdU or other thymidine analogues when testing various administration routes or protocols.
authors.affiliation: Institute of Biology and Ecology, Faculty of Science, P. J. Šafárik University in Košice, Košice, Slovakia. juraj.sevc@upjs.sk.
authors.forename: J
authors.initials: J
authors.lastname: Mikeš
authors.forename: J
authors.initials: J
authors.lastname: Ševc
authors.forename: J
authors.initials: J
authors.lastname: Košuth
authors.forename: A
authors.initials: A
authors.lastname: Matiašová
authors.forename: Z
authors.initials: Z
authors.lastname: Daxnerová
authors.forename: P
authors.initials: P
authors.lastname: Fedoročko
mesh_terms: D000818:Animals; D000963:Antimetabolites / Q000097:blood*; D001973:Bromodeoxyuridine / Q000097:blood*; D002448:Cell Adhesion; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D005434:Flow Cytometry; D006801:Humans; D007274:Injections, Intraperitoneal; D051381:Rats
chemical_list: D000963:Antimetabolites; D001973:Bromodeoxyuridine