title: Enrichment for chemoresistant ovarian cancer stem cells from human cell lines.
abstract: Cancer stem cells (CSCs) are defined as a subset of slow cycling and undifferentiated cells that divide asymmetrically to generate highly proliferative, invasive, and chemoresistant tumor cells. Therefore, CSCs are an attractive population of cells to target therapeutically. CSCs are predicted to contribute to a number of types of malignancies including those in the blood, brain, lung, gastrointestinal tract, prostate, and ovary. Isolating and enriching a tumor cell population for CSCs will enable researchers to study the properties, genetics, and therapeutic response of CSCs. We generated a protocol that reproducibly enriches for ovarian cancer CSCs from ovarian cancer cell lines (SKOV3 and OVCA429). Cell lines are treated with 20 µM cisplatin for 3 days. Surviving cells are isolated and cultured in a serum-free stem cell media containing cytokines and growth factors. We demonstrate an enrichment of these purified CSCs by analyzing the isolated cells for known stem cell markers Oct4, Nanog, and Prom1 (CD133) and cell surface expression of CD177 and CD133. The CSCs exhibit increased chemoresistance. This method for isolation of CSCs is a useful tool for studying the role of CSCs in chemoresistance and tumor relapse.
authors.affiliation: Department of Biochemistry and Molecular Biology, Indiana University School of Medicine.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Cole
authors.affiliation: Department of Biochemistry and Molecular Biology, Indiana University School of Medicine.
authors.forename: Stancy
authors.initials: S
authors.lastname: Joseph
authors.affiliation: Department of Biochemistry and Molecular Biology, Indiana University School of Medicine.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Sudhahar
authors.affiliation: Department of Biochemistry and Molecular Biology, Indiana University School of Medicine; kcowdend@iupui.edu.
authors.forename: Karen D
authors.initials: KD
authors.lastname: Cowden Dahl
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D014408:Biomarkers, Tumor / Q000378:metabolism; D000077216:Carcinoma, Ovarian Epithelial; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D016895:Culture Media, Serum-Free; D019008:Drug Resistance, Neoplasm; D005260:Female; D006801:Humans; D009375:Neoplasms, Glandular and Epithelial / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D018874:Spheroids, Cellular
chemical_list: D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D016895:Culture Media, Serum-Free; D002945:Cisplatin