title: Current approaches in identification and isolation of human renal cell carcinoma cancer stem cells.
abstract: In recent years, cancer stem cells (CSCs)/tumor initiating cells (TICs) have been identified inside different tumors. However, currently used anti-cancer therapies are mostly directed against somatic tumor cells without targeting CSCs/TICs. CSCs/TICs also gain resistance to chemotherapies/radiotherapies. For the development of efficient treatment strategies, choosing the best method for isolation and characterization of CSCs/TICs is still debated among the scientific community. In this review, we summarize recent data concerning isolation techniques for CSCs using magnetic cell sorting and flow cytometry. The review focuses on the strategies for sample preparation during flow cytometric analysis, elaborating biomarkers such as CXCR4, CD105, and CD133. In addition, functional properties characteristic of CSCs/TICs using side population selection through Hoechst 33342 dye, aldehyde dehydrogenase 1, dye-cycle violet, and rhodamine 123 are also discussed. We also include a special focus on enriching CSCs/TICs using three-dimensional cell culture models such as agarose-agarose microbeads and sphere formation.
authors.affiliation: Molecular Oncology Laboratory, Clinic of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland. imrankhanbioinfo@gmail.com.
authors.forename: Mohammed I
authors.initials: MI
authors.lastname: Khan
authors.affiliation: Molecular Oncology Laboratory, Clinic of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Czarnecka
authors.affiliation: Molecular Oncology Laboratory, Clinic of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland.
authors.forename: Igor
authors.initials: I
authors.lastname: Helbrecht
authors.affiliation: Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, 02-106, Warsaw, Poland.|Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02-106, Warsaw, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Bartnik
authors.affiliation: Department of Urology, Emory University School of Medicine, Atlanta, GA, 30322, USA.
authors.forename: Fei
authors.initials: F
authors.lastname: Lian
authors.affiliation: Molecular Oncology Laboratory, Clinic of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141, Warsaw, Poland.
authors.forename: Cezary
authors.initials: C
authors.lastname: Szczylik
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D002292:Carcinoma, Renal Cell / Q000473:pathology*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D014411:Neoplastic Stem Cells / Q000145:classification / Q000166:cytology* / Q000378:metabolism; D061251:Primary Cell Culture / Q000379:methods*
chemical_list: D014408:Biomarkers, Tumor