title: Cellular characteristics of head and neck cancer stem cells in type IV collagen-coated adherent cultures.
abstract: Although head and neck squamous carcinoma cancer stem cells (HNSC-CSCs) can be enriched in serum-free suspension cultures, it is difficult to stably expand HNSC-CSC lines in suspension due to spontaneous apoptosis and differentiation. Here, we investigated whether HNSC-CSCs can be expanded without loss of stem cell properties by adherent culture methods. Cell culture plates were coated with type IV collagen, laminin, or fibronectin. We examined cancer stem cell traits of adherent HNSC-CSCs grown on these plates using immunocytochemistry for stem cell marker expression and analyses of chemo-resistance and xenograft tumorigenicity. We also assessed the growth rate, apoptosis rate, and gene transduction efficiency of adherent and suspended HNSC-CSCs. HNSC-CSCs grew much faster on type IV collagen-coated plates than in suspension. Adherent HNSC-CSCs expressed putative stem cell markers (OCT4 and CD44) and were chemo-resistant to various cytotoxic drugs (cisplatin, fluorouracil, paclitaxel, and docetaxel). Adherent HNSC-CSCs at the limiting dilution (1000 cells) produced tumors in nude mice. Adherent HNSC-CSCs also showed less spontaneous apoptotic cell death and were more competent to lentiviral transduction than suspended HNSC-CSCs. In conclusion, compared to suspension cultures, adherence on type IV collagen-coated culture plates provides better experimental conditions for HNSC-CSC expansion, which should facilitate various refined cellular studies.
authors.affiliation: Department of Otorhinolaryngology-Head and Neck Surgery, Research Institute of Medical Science, Konkuk University School of Medicine, Republic of Korea.
authors.forename: Young Chang
authors.initials: YC
authors.lastname: Lim
authors.forename: Se-Yeong
authors.initials: SY
authors.lastname: Oh
authors.forename: Hyunggee
authors.initials: H
authors.lastname: Kim
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology; D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D048430:Cell Shape; D002470:Cell Survival; D024141:Collagen Type IV / Q000737:chemistry*; D019008:Drug Resistance, Neoplasm; D005260:Female; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D006258:Head and Neck Neoplasms / Q000473:pathology*; D006801:Humans; D055767:Immobilized Proteins; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D010641:Phenotype; D011994:Recombinant Proteins / Q000096:biosynthesis; D018874:Spheroids, Cellular / Q000502:physiology; D014162:Transfection
chemical_list: D000943:Antigens, Differentiation; D000970:Antineoplastic Agents; D024141:Collagen Type IV; D055767:Immobilized Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins