title: Transfer printing of transfected cell microarrays from poly(ethylene glycol)-oleyl surfaces onto biological hydrogels.
abstract: We have developed a novel technique for constructing microarrays of transfected mammalian cells on or in extracellular matrix (ECM) hydrogels by transfer printing from patterned poly(ethylene glycol) (PEG)-oleyl surfaces. A mixed solution of small interfering RNA (siRNA) and a transfection reagent was spotted on PEG-oleyl-coated glass slides using an ink-jet printer, and the cells were then transiently immobilized on the patterned transfection mixtures. After overlaying an ECM hydrogel sheet onto the immobilized cells, the cells sandwiched between the glass slide and the hydrogel sheet were incubated at 37°C for simultaneous transfection of siRNA into cells and adhesion of cells to the hydrogel sheet. Transfer of the adhered, transfected cells was completed by peeling off the hydrogel sheet. The knockdown of a model gene in the transferred cell microarray by the transfected siRNA was successfully confirmed. Transfected cell microarrays were also embedded within three-dimensional ECM hydrogels. In the three-dimensional hydrogel, the inhibition effect of siRNA on cancer cell invasion was evaluated by quantifying the size of cell clusters on the microarrays. These results indicate that transfection of cell microarrays on or in a biological matrix is a promising technique for high-throughput screening of disease-related genes by direct observation of cellular phenomena in a physiologically relevant environment.
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yamaguchi
authors.forename: Senori
authors.initials: S
authors.lastname: Komiya
authors.forename: Erika
authors.initials: E
authors.lastname: Matsunuma
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamahira
authors.forename: Takanori
authors.initials: T
authors.lastname: Kihara
authors.forename: Jun
authors.initials: J
authors.lastname: Miyake
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Nagamune
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D019855:Ethylene Glycol; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D020100:Hydrogels; D008322:Mammals; D046228:Microarray Analysis / Q000379:methods*
chemical_list: D020100:Hydrogels; D019855:Ethylene Glycol
keywords: cell invasion; cell microarray; cell printing; extracellular matrix; phagocytosis; reverse transfection