title: Transfecting Human Monocytes with RNA.
abstract: Targeting monocytes as a delivery system for drugs or nucleic acids, and thereby harnessing their natural tissue-infiltrating capacity, has become an area of intense investigation in both basic and clinical research. Herein we describe an efficient method to deliver mRNA (messenger RNA) or siRNA (small interfering RNA) into human monocytes by electroporation. This method can be applied in the laboratory to monocytes isolated via magnetic bead-based techniques, or in a clinical setting using monocytes that were collected via counterflow centrifugation elutriation using the Elutra(®) Cell Separation System. We further demonstrate that electroporation of monocytes with RNA represents a robust and highly relevant approach to modify monocytes for cell-based therapies. Last, the procedure described can readily be adapted to monocytes from different species, hence facilitating research in animal models.
authors.affiliation: Department of Surgery, Duke University School of Medicine, Durham, NC, 27710, USA.
authors.forename: Jens
authors.initials: J
authors.lastname: Dannull
authors.affiliation: Department of Surgery, Duke University School of Medicine, Durham, NC, 27710, USA. smita.nair@duke.edu.
authors.forename: Smita K
authors.initials: SK
authors.lastname: Nair
mesh_terms: D002469:Cell Separation; D002478:Cells, Cultured; D055590:Electromagnetic Phenomena; D018274:Electroporation / Q000379:methods*; D006801:Humans; D009000:Monocytes / Q000166:cytology* / Q000378:metabolism; D012313:RNA / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012313:RNA
keywords: Electroporation; Elutriation; Monocytes; RNA transfection; siRNA transfection