Isolation of cancer stem cells from human prostate cancer samples

Samuel J. Vidal, S. Aidan Quinn, Janis de la Iglesia-Vicente, Dennis M. Bonal, Veronica Rodriguez-Bravo, Adolfo Firpo-Betancourt, Carlos Cordon-Cardo, Josep Domingo-Domenech

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish (US)
Article numbere51332
JournalJournal of Visualized Experiments
Issue number85
StatePublished - Mar 14 2014


  • Cancer Stem Cells
  • Castration Resistant Prostate Cancer
  • HLA class I
  • Human Tissue Samples
  • Intratumoral heterogeneity
  • Issue 85
  • Medicine
  • Metastatic Prostate Cancer
  • Primary Prostate Cancer
  • Prostate Cancer
  • Tumor Initiating Cells

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology


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