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Nat Protoc. 2009;4(9):1313-27. doi: 10.1038/nprot.2009.117. Epub 2009 Aug 20.

Measurement of proliferation and disappearance of rapid turnover cell populations in human studies using deuterium-labeled glucose.

Author information

1
Centre for Infection, Cellular and Molecular Medicine, St George's, University of London, London, UK. macallan@sgul.ac.uk

Abstract

Cell proliferation may be measured in vivo by quantifying DNA synthesis with isotopically labeled deoxyribonucleotide precursors. Deuterium-labeled glucose is one such precursor which, because it achieves high levels of enrichment for a short period, is well suited to the study of rapidly dividing cells, in contrast to the longer term labeling achieved with heavy water ((2)H(2)O). As deuterium is non-radioactive and glucose can be readily administered, this approach is suitable for clinical studies. It has been widely applied to investigate human lymphocyte proliferation, but solid tissue samples may also be analyzed. Rate, duration and route (intravenous or oral) of [6,6-(2)H(2)]-glucose administration should be adapted to the target cell of interest. For lymphocytes, cell separation is best achieved by fluorescence activated cell sorting (FACS), although magnetic bead separation is an alternative. DNA is then extracted, hydrolyzed enzymatically and analyzed by gas chromatography mass spectrometry (GC/MS). Appropriate mathematical modeling is critical to interpretation. Typical time requirements are as follows: labeling, 10-24 h; sampling, approximately 3 weeks; DNA extraction/derivatization, 2-3 d; and GC/MS analysis, approximately 2 d.

PMID:
19696750
DOI:
10.1038/nprot.2009.117
[Indexed for MEDLINE]

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