title: Dielectrophoresis-assisted 3D nanoelectroporation for non-viral cell transfection in adoptive immunotherapy.
abstract: Current transfection technologies lead to significant inter-clonal variations. Previously we introduced a unique electrotransfection technology, Nanochannel-Electroporation (NEP), which can precisely and benignly transfect small cell populations (~100-200 cells) with single-cell resolution. Here we report on the development of a novel 3D NEP system for large scale transfection. A properly-engineered array of nanochannels, capable of handling/transfecting ~60 000 cells cm(-2), was fabricated using cleanroom technologies. Positive dielectrophoresis was used to selectively position cells on the nanochannels, thus allowing highly efficient transfection. Single-cell dosage control was demonstrated using both small and large molecules, and different cell types. The potential clinical relevance of this system was tested with difficult-to-transfect natural killer cell suspensions, and plasmids encoding for the chimeric antigen receptor (CAR), a model of high relevance for adoptive immunotherapy. Our results show significantly higher CAR transfection efficiencies for the DEP-NEP system (>70% vs. <30%), as well as enhanced cell viabilities.
authors.affiliation: Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio 43210, USA. lee.31@osu.edu.
authors.forename: Lingqian
authors.initials: L
authors.lastname: Chang
authors.forename: Daniel
authors.initials: D
authors.lastname: Gallego-Perez
authors.forename: Xi
authors.initials: X
authors.lastname: Zhao
authors.forename: Paul
authors.initials: P
authors.lastname: Bertani
authors.forename: Zhaogang
authors.initials: Z
authors.lastname: Yang
authors.forename: Chi-Ling
authors.initials: CL
authors.lastname: Chiang
authors.forename: Veysi
authors.initials: V
authors.lastname: Malkoc
authors.forename: Junfeng
authors.initials: J
authors.lastname: Shi
authors.forename: Chandan K
authors.initials: CK
authors.lastname: Sen
authors.forename: Lynn
authors.initials: L
authors.lastname: Odonnell
authors.forename: Jianhua
authors.initials: J
authors.lastname: Yu
authors.forename: Wu
authors.initials: W
authors.lastname: Lu
authors.forename: L James
authors.initials: LJ
authors.lastname: Lee
mesh_terms: D000818:Animals; D002460:Cell Line; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000295:instrumentation / Q000379:methods*; D007694:Killer Cells, Natural / Q000166:cytology; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods*; D051381:Rats; D014162:Transfection / Q000295:instrumentation / Q000379:methods*; D019076:Transgenes