title: Isolation of cancer stem cells from human prostate cancer samples.
abstract: The cancer stem cell (CSC) model has been considerably revisited over the last two decades. During this time CSCs have been identified and directly isolated from human tissues and serially propagated in immunodeficient mice, typically through antibody labeling of subpopulations of cells and fractionation by flow cytometry. However, the unique clinical features of prostate cancer have considerably limited the study of prostate CSCs from fresh human tumor samples. We recently reported the isolation of prostate CSCs directly from human tissues by virtue of their HLA class I (HLAI)-negative phenotype. Prostate cancer cells are harvested from surgical specimens and mechanically dissociated. A cell suspension is generated and labeled with fluorescently conjugated HLAI and stromal antibodies. Subpopulations of HLAI-negative cells are finally isolated using a flow cytometer. The principal limitation of this protocol is the frequently microscopic and multifocal nature of primary cancer in prostatectomy specimens. Nonetheless, isolated live prostate CSCs are suitable for molecular characterization and functional validation by transplantation in immunodeficient mice.
authors.affiliation: Department of Pathology, Icahn School of Medicine at Mount Sinai.
authors.forename: Samuel J
authors.initials: SJ
authors.lastname: Vidal
authors.affiliation: Department of Pathology, Icahn School of Medicine at Mount Sinai.
authors.forename: S Aidan
authors.initials: SA
authors.lastname: Quinn
authors.affiliation: Department of Pathology, Icahn School of Medicine at Mount Sinai.
authors.forename: Janis
authors.initials: J
authors.lastname: de la Iglesia-Vicente
authors.affiliation: Department of Pathology, Icahn School of Medicine at Mount Sinai.
authors.forename: Dennis M
authors.initials: DM
authors.lastname: Bonal
authors.affiliation: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center.
authors.forename: Veronica
authors.initials: V
authors.lastname: Rodriguez-Bravo
authors.affiliation: Department of Pathology, Icahn School of Medicine at Mount Sinai.
authors.forename: Adolfo
authors.initials: A
authors.lastname: Firpo-Betancourt
authors.affiliation: Department of Pathology, Icahn School of Medicine at Mount Sinai.
authors.forename: Carlos
authors.initials: C
authors.lastname: Cordon-Cardo
authors.affiliation: Department of Pathology, Icahn School of Medicine at Mount Sinai; josep.domingo-domenech@mssm.edu.
authors.forename: Josep
authors.initials: J
authors.lastname: Domingo-Domenech
mesh_terms: D003584:Cytological Techniques / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008297:Male; D014411:Neoplastic Stem Cells / Q000737:chemistry / Q000473:pathology*; D011471:Prostatic Neoplasms / Q000737:chemistry / Q000473:pathology*; D064129:Prostatic Neoplasms, Castration-Resistant / Q000737:chemistry / Q000473:pathology; D013194:Staining and Labeling / Q000379:methods