title: Magselectofection: an integrated method of nanomagnetic separation and genetic modification of target cells.
abstract: Research applications and cell therapies involving genetically modified cells require reliable, standardized, and cost-effective methods for cell manipulation. We report a novel nanomagnetic method for integrated cell separation and gene delivery. Gene vectors associated with magnetic nanoparticles are used to transfect/transduce target cells while being passaged and separated through a high gradient magnetic field cell separation column. The integrated method yields excellent target cell purity and recovery. Nonviral and lentiviral magselectofection is efficient and highly specific for the target cell population as demonstrated with a K562/Jurkat T-cell mixture. Both mouse and human enriched hematopoietic stem cell pools were effectively transduced by lentiviral magselectofection, which did not affect the hematopoietic progenitor cell number determined by in vitro colony assays. Highly effective reconstitution of T and B lymphocytes was achieved by magselectofected murine wild-type lineage-negative Sca-1(+) cells transplanted into Il2rg(-/-) mice, stably expressing GFP in erythroid, myeloid, T-, and B-cell lineages. Furthermore, nonviral, lentiviral, and adenoviral magselectofection yielded high transfection/transduction efficiency in human umbilical cord mesenchymal stem cells and was fully compatible with their differentiation potential. Upscaling to a clinically approved automated cell separation device was feasible. Hence, once optimized, validated, and approved, the method may greatly facilitate the generation of genetically engineered cells for cell therapies.
authors.affiliation: Institute of Experimental Oncology and Therapy Research, Technische Universität München, Munich, Germany.
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Sanchez-Antequera
authors.forename: Olga
authors.initials: O
authors.lastname: Mykhaylyk
authors.forename: Niek P
authors.initials: NP
authors.lastname: van Til
authors.forename: Arzu
authors.initials: A
authors.lastname: Cengizeroglu
authors.forename: J Henk
authors.initials: JH
authors.lastname: de Jong
authors.forename: Marshall W
authors.initials: MW
authors.lastname: Huston
authors.forename: Martina
authors.initials: M
authors.lastname: Anton
authors.forename: Ian C D
authors.initials: IC
authors.lastname: Johnston
authors.forename: Zygmunt
authors.initials: Z
authors.lastname: Pojda
authors.forename: Gerard
authors.initials: G
authors.lastname: Wagemaker
authors.forename: Christian
authors.initials: C
authors.lastname: Plank
mesh_terms: D000818:Animals; D000950:Antigens, Ly / Q000235:genetics; D002469:Cell Separation / Q000379:methods*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000737:chemistry; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D053631:Interleukin Receptor Common gamma Subunit / Q000235:genetics; D019169:Jurkat Cells; D020014:K562 Cells; D008280:Magnetics; D008565:Membrane Proteins / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry; D014162:Transfection
chemical_list: D000950:Antigens, Ly; C507942:Il2rg protein, mouse; D053631:Interleukin Receptor Common gamma Subunit; C074650:Ly6a protein, mouse; D008565:Membrane Proteins