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  • How reliable are sino-nasal cell lines for studying the pathophysiology of chronic rhinosinusitis?

How reliable are sino-nasal cell lines for studying the pathophysiology of chronic rhinosinusitis?

The Annals of otology, rhinology, and laryngology (2014-12-30)
Stephen L Ball, Monika I Suwara, Lee A Borthwick, Janet A Wilson, Derek A Mann, Andrew J Fisher
초록

Well-characterized cell lines represent useful scientific tools to study the pathophysiology of human disease. Chronic rhinosinusitis (CRS) is a very common condition, though the number of CRS cell lines is limited, as are data showing how closely they resemble primary cells. Searches for available human cell lines were performed using the American Type Culture Collection (ATCC) and European Collection of Cell Cultures (ECACC). Identified cells were cultured and characterized with tinctorial and immunohistochemical staining and ELISA to assess their response to common, disease-relevant inflammatory stimuli. Carefully phenotyped CRS patients were recruited with informed consent. Primary nasal epithelial cell (PNEC) brushings were harvested, cultured, and compared to the available cell lines. Searches identified 1 relevant CRS sino-nasal cell line, RPMI 2650. Cultured PNECs showed strong expression of epithelial markers while being negative for mesenchymal markers. However, RPMI 2650 cells show an atypical mixed epithelial/mesenchymal phenotype. When stimulated by pro-inflammatory ligands, PNECs responded in a dose-dependent manner, whereas RPMI 2650 cells showed limited response. The number and availability of cell lines to study the pathophysiology of CRS greatly underrepresent the disease burden. Additionally, the sole commercially available cell line appears to have a different phenotype and behavior to primary patient-derived cells. The development of further reproducible cell lines would be beneficial in our understanding of CRS.

MATERIALS
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Polyinosinic–polycytidylic acid potassium salt, with buffer salts, TLR ligand tested
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TWEEN® 20, viscous liquid
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Lipopolysaccharides from Escherichia coli O111:B4, purified by phenol extraction
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L-Glutamine solution, 200 mM, solution, sterile-filtered, BioXtra, suitable for cell culture
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Penicillin-Streptomycin, with 10,000 units penicillin and 10 mg streptomycin per mL in 0.9% NaCl, 0.1 μm filtered, BioReagent, suitable for cell culture
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L-Glutamine
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L-Glutamine, BioUltra, ≥99.5% (NT)
SAFC
L-Glutamine
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Anti-Rabbit IgG (whole molecule)–TRITC antibody produced in goat, affinity isolated antibody, buffered aqueous solution
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L-Glutamine, γ-irradiated, BioXtra, suitable for cell culture
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L-Glutamine, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
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Triton X-100, laboratory grade
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Minimum Essential Medium Eagle, With Earle′s salts and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
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Triton X-100, for molecular biology
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L-Glutamine, ReagentPlus®, ≥99% (HPLC)
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ECO TWEEN® 20, viscous liquid