title: Measurement of cell proliferation by heavy water labeling.
abstract: DNA replication occurs almost exclusively during S-phase of the cell cycle and represents a simple biochemical metric of cell division. Previous methods for measuring cell proliferation rates have important limitations. Here, we describe experimental protocols for measuring cell proliferation and death rates based on the incorporation of deuterium ((2)H) from heavy water ((2)H(2)O) into the deoxyribose moiety of purine deoxyribonucleotides in DNA of dividing cells. Label incorporation is measured by gas chromatography/mass spectrometry. Modifications of the basic protocol permit analysis of small cell samples (down to 2,000 cells). The theoretical basis and operational requirements for effective use of these methods to measure proliferation and death rates of cells in vivo are described. These methods are safe for use in humans, have technical and interpretation advantages over alternative techniques and can be used on small numbers of cells. The protocols enable definitive in vivo studies of the fraction or absolute number of newly divided cells and their subsequent survival kinetics in animals and humans.
authors: KineMed Inc., 5980 Horton Street, Emeryville, California 94608, USA.
authors: Robert
authors: R
authors: Busch
authors: Richard A
authors: RA
authors: Neese
authors: Mohamad
authors: M
authors: Awada
authors: Gregory M
authors: GM
authors: Hayes
authors: Marc K
authors: MK
authors: Hellerstein
mesh_terms: D000818:Animals; D001834:Body Water; D016923:Cell Death; D049109:Cell Proliferation; D003106:Colon / Q000166:cytology; D004261:DNA Replication; D017666:Deuterium Oxide / Q000032:analysis* / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology; D006801:Humans; D007553:Isotope Labeling; D015603:Keratinocytes / Q000166:cytology; D051379:Mice; D032342:Muscle Cells / Q000166:cytology; D009369:Neoplasms; D009474:Neurons / Q000166:cytology; D051381:Rats; D046509:Tissue Culture Techniques
chemical_list: D017666:Deuterium Oxide