title: Silver nanoparticles inhibit fish gill cell proliferation in protein-free culture medium.
abstract: While short-term exposures of vertebrate cells, such as from fish, can be performed in defined, serum-free media, long-term cultures generally require addition of growth factors and proteins, normally supplied with a serum supplement. However, proteins are known to alter nanoparticle properties by binding to nanoparticles. Therefore, in order to be able to study nanoparticle-cell interactions for extended periods, the rainbow trout (Oncorhynchus mykiss) gill cell line, RTgill-W1, was adapted to proliferate in a commercial, serum-free medium, InVitrus VP-6. The newly adapted cell strain was named RTgill-W1-pf (protein free). These cells proliferate at a speed similar to the RTgill-W1 cells cultured in a fully supplemented medium containing 5% fetal bovine serum. As well, they were successfully cryopreserved in liquid nitrogen and fully recovered after thawing. Yet, senescence set in after about 10 passages in InVitrus VP-6 medium, revealing that this medium cannot fully support long-term culture of the RTgill-W1 strain. The RTgill-W1-pf cell line was subsequently applied to investigate the effect of silver nanoparticles (AgNP) on cell proliferation over a period of 12 days. Indeed, cell proliferation was inhibited by 10 μM AgNP. This effect correlated with high levels of silver being associated with the cells. The new cell line, RTgill-W1-pf, can serve as a unique representation of the gill cell-environment interface, offering novel opportunities to study nanoparticle-cell interactions without serum protein interference.
authors.affiliation: a Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology , Dübendorf , Switzerland .|b École Polytechnique Fédérale de Lausanne, School of Architecture, Civil and Environmental Engineering , Lausanne , Switzerland , and.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yue
authors.affiliation: a Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology , Dübendorf , Switzerland .|c ETH (EidgenÖssische Technische Hochschule) Zürich, Department of Environmental Systems Sciences , Zürich , Switzerland.
authors.forename: Renata
authors.initials: R
authors.lastname: Behra
authors.affiliation: a Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology , Dübendorf , Switzerland .|c ETH (EidgenÖssische Technische Hochschule) Zürich, Department of Environmental Systems Sciences , Zürich , Switzerland.
authors.forename: Laura
authors.initials: L
authors.lastname: Sigg
authors.affiliation: a Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Toxicology , Dübendorf , Switzerland .|b École Polytechnique Fédérale de Lausanne, School of Architecture, Civil and Environmental Engineering , Lausanne , Switzerland , and.|c ETH (EidgenÖssische Technische Hochschule) Zürich, Department of Environmental Systems Sciences , Zürich , Switzerland.
authors.forename: Kristin
authors.initials: K
authors.lastname: Schirmer
mesh_terms: D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free; D005880:Gills / Q000166:cytology / Q000187:drug effects*; D053768:Metal Nanoparticles / Q000737:chemistry / Q000633:toxicity*; D017686:Oncorhynchus mykiss; D012834:Silver / Q000737:chemistry / Q000633:toxicity*
chemical_list: D016895:Culture Media, Serum-Free; D012834:Silver
keywords: Gill; long-term exposure; rainbow trout; serum-free medium; silver nanoparticles