TY - JOUR
T1 - Cellular characteristics of head and neck cancer stem cells in type IV collagen-coated adherent cultures
AU - Lim, Young Chang
AU - Oh, Se Yeong
AU - Kim, Hyunggee
N1 - Funding Information:
This study was supported by grants from the National Research Foundation of Korea (NRF) funded by the Korean government (MEST) (No. 2010-0022256 to Y.C. Lim) and from the National R&D Program for Cancer Control, Ministry for Health and Welfare , the Republic of Korea (No. 2008-0058785 to H. Kim).
PY - 2012/6/10
Y1 - 2012/6/10
N2 - 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.
AB - 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.
KW - Adherent cultures
KW - Cancer stem cell
KW - Head neck cancer
KW - Type IV collagen
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UR - http://www.scopus.com/inward/citedby.url?scp=84862813687&partnerID=8YFLogxK
U2 - 10.1016/j.yexcr.2012.02.038
DO - 10.1016/j.yexcr.2012.02.038
M3 - Article
C2 - 22426199
AN - SCOPUS:84862813687
VL - 318
SP - 1104
EP - 1111
JO - Experimental Cell Research
JF - Experimental Cell Research
SN - 0014-4827
IS - 10
ER -
