Flow cytometric detection of prostate tumor cells using chemoaffinity labels

The Prostate
Lisa Y WuClifford E Berkman

Abstract

The enzyme-biomarker prostate-specific membrane antigen (PSMA) is an emerging target for imaging and therapeutic applications for prostate cancer. However, the use of PSMA for detecting circulating prostate tumor cells remains under-explored. The present study focuses on the specific labeling of PSMA+ prostate cancer cells with a fluorescent PSMA inhibitor and the quantitation of PSMA+ cells in blood by flow cytometry (FC) using a gating strategy to separate labeled PSMA+ cells from peripheral blood mononuclear cells. Suspensions of PSMA+ (LNCaP) and PSMA- (DU145) cells were incubated with the fluorescent PSMA inhibitor FAMX-CTT-54. Incubation parameters (time, temperature, and label concentration) were varied to optimize cell labeling. A gating protocol based on double fluorescent labeling of CD45 and PSMA was developed for the quantitiation of LNCaP cells in the presence of white blood cells from bovine blood. Nonfluorescent beads were added to the labeled cell mixture and served as internal standard for precise cellular quantification of LNCaP cells by flow cytometry. The fluorescent PSMA inhibitor FAMX-CTT-54 was specific for PSMA+ cells. The minimum time and concentration of FAMX-CTT-54 for effective labeling of PSMA+ cell...Continue Reading

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Citations

Dec 17, 2011·Nutrition and Cancer·Ajit Vikram, Gopabandhu Jena
May 9, 2014·Molecular Diagnosis & Therapy·Giuseppe GallettiDavid M Nanus
Jun 29, 2011·Expert Review of Molecular Diagnostics·Marco DanovaGiuliano Mazzini
Apr 11, 2012·The Prostate·Lisa Y WuClifford E Berkman
Apr 24, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Kai PangZhengqin Gu

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