Cellular characteristics of head and neck cancer stem cells in type IV collagen-coated adherent cultures

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 l...

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Bibliographic Details
Published in:Experimental cell research 2012-06, Vol.318 (10), p.1104-1111
Main Authors: Lim, Young Chang, Oh, Se-Yeong, Kim, Hyunggee
Format: Article
Language:English
Subjects:
Adherent cultures
Animals
Antigens, Differentiation - genetics
Antigens, Differentiation - metabolism
Antineoplastic Agents - pharmacology
Cancer stem cell
Carcinoma, Squamous Cell - pathology
Cell Adhesion
Cell culture
Cell Culture Techniques
Cell Differentiation
Cell Proliferation
Cell Shape
Cell Survival
Cellular biology
Collagen Type IV - chemistry
Drug Resistance, Neoplasm
Female
Green Fluorescent Proteins - biosynthesis
Head & neck cancer
Head and Neck Neoplasms - pathology
Humans
Immobilized Proteins
Mice
Mice, Nude
Neoplasm Transplantation
Neoplastic Stem Cells - drug effects
Neoplastic Stem Cells - metabolism
Neoplastic Stem Cells - physiology
Phenotype
Recombinant Proteins - biosynthesis
Research methodology
Spheroids, Cellular - physiology
Stem cells
Transfection
Type IV collagen
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Summary: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. ► Head and neck cancer stem cell (HNCSC) can be enriched by suspension cultures ► It is difficult to stably expand HNCSC as a floating sphere-form in vitro ► We invented HNCSC expansion by adherent culture methods using type IV collagen. ► Our study provides a significant advantage to study HNCSC biology.
ISSN:0014-4827
1090-2422
DOI:10.1016/j.yexcr.2012.02.038