Flow cytometric method for estimation of 5-bromo-2'deosyuridine content in rat serum

Title:

Flow cytometric method for estimation of 5-bromo-2'deosyuridine content in rat serum

Creator:

Jaromír Mikeš, Ševc, J., Košuth, J., Matiašová, A., Zuzana Daxnerová, and Peter Fedoročko

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:f27f69bb-a282-40e7-a789-8f8e4b5c6e48
uuid:f27f69bb-a282-40e7-a789-8f8e4b5c6e48
issn:0862-8408

Subject:

Fyziologie člověka a srovnávací fyziologie, proliferace buněk, průtoková cytometrie, cell proliferation, flow cytometry, BrdU pharmacokinetic, BrdU incorporation, HCT-116, 14, and 612

Type:

article, články, model:article, and TEXT

Format:

print, bez média, and svazek

Description:

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 μmol.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., J. Mikeš, J. Ševc, J. Košuth, A. Matiašová, Z. Daxnerová, P. Fedoročko., and Obsahuje bibliografii

Language:

English

Rights:

http://creativecommons.org/licenses/by-nc-sa/4.0/
policy:public

Source:

Physiological research | 2014 Volume:63 | Number:6

Harvested from:

CDK

Metadata only:

false