Identification of resting cells by dual-parameter flow cytometry of statin expression and DNA content

Cytometry
C PellicciariE Wang

Abstract

Statin, a 57-kDa nuclear protein, has been recognized as a unique marker of quiescent (G0) cells; specific monoclonal antibodies (MoAb) against statin have been produced and used to label resting cells in tissue sections and in cultured cells. We present an improved method for the identification of G0 cells by dual-parameter flow cytometry of statin expression and DNA content. The appropriate technical conditions were set up by using resting and cycling human fibroblasts as a model cell system. Several fixatives proved to be suitable for the immunocytochemical detection of statin; among them, 70% ethanol was selected because this fixation procedure is suitable for DNA staining with intercalating dyes and is routinely used for the immunolabeling of proliferation markers (such as proliferating cell nuclear antigen [PCNA] and Ki-67) and of bromodeoxyuridine (BrdUrd) incorporation. Following cell permeabilization with detergent, exposure to the antistatin antibody (S-44), and indirect fluorescein isothiocyanate immunolabeling, cells were counterstained for DNA with propidium iodide and analyzed by dual-parameter flow cytometry. In cells from several animal sources (rat thymocytes and C6 glioma cells, mouse 3T3 cells, and human MCF-...Continue Reading

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Citations

Aug 13, 2002·Cloning and Stem Cells·Björn Oback, David Wells
Sep 12, 2013·Cellular Reprogramming·D N Wells
Dec 15, 2006·The Journal of Gene Medicine·Hsiao-Ping LeeYu-Chen Hu
May 16, 2006·Mathematical Biosciences·Lorenzo SpinelliBritta Basse
Aug 1, 2002·Oncogene·Ana M TariGabriel Lopez-Berestein
Sep 28, 2001·Hematological Oncology·H G Jørgensen, T L Holyoake

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