title: High-Throughput Mechanobiology Screening Platform Using Micro- and Nanotopography.
abstract: We herein demonstrate the first 96-well plate platform to screen effects of micro- and nanotopographies on cell growth and proliferation. Existing high-throughput platforms test a limited number of factors and are not fully compatible with multiple types of testing and assays. This platform is compatible with high-throughput liquid handling, high-resolution imaging, and all multiwell plate-based instrumentation. We use the platform to screen for topographies and drug-topography combinations that have short- and long-term effects on T cell activation and proliferation. We coated nanofabricated "trench-grid" surfaces with anti-CD3 and anti-CD28 antibodies to activate T cells and assayed for interleukin 2 (IL-2) cytokine production. IL-2 secretion was enhanced at 200 nm trench width and >2.3 μm grating pitch; however, the secretion was suppressed at 100 nm width and <0.5 μm pitch. The enhancement on 200 nm grid trench was further amplified with the addition of blebbistatin to reduce contractility. The 200 nm grid pattern was found to triple the number of T cells in long-term expansion, a result with direct clinical applicability in adoptive immunotherapy.
authors.forename: Junqiang
authors.initials: J
authors.lastname: Hu
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Gondarenko
authors.forename: Alex P
authors.initials: AP
authors.lastname: Dang
authors.forename: Keenan T
authors.initials: KT
authors.lastname: Bashour
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
authors.forename: Roddy S
authors.initials: RS
authors.lastname: O'Connor
authors.forename: Sunwoo
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Pathology, New York University School of Medicine , New York, New York 10016, United States.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Liapis
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
authors.forename: Saba
authors.initials: S
authors.lastname: Ghassemi
authors.affiliation: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Milone
authors.forename: Michael P
authors.initials: MP
authors.lastname: Sheetz
authors.affiliation: Nuffield Department of Orthopedics, Rheumatology and Musculoskeletal Sciences, Kennedy Institute of Rheumatology, The University of Oxford , Oxford, OX3 7FY, U.K.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Dustin
authors.forename: Lance C
authors.initials: LC
authors.lastname: Kam
authors.forename: James C
authors.initials: JC
authors.lastname: Hone
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D008213:Lymphocyte Activation; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism
chemical_list: C508594:IL2 protein, human; D007376:Interleukin-2
keywords: High-throughput screening; Lck; T cell; interleukin-2; long-term expansion; nanotechnology