title: Magnetoelectroporation: improved labeling of neural stem cells and leukocytes for cellular magnetic resonance imaging using a single FDA-approved agent.
abstract: Cellular magnetic resonance imaging (MRI) relies on the use of intracellular contrast agents, primarily iron oxide compounds. Several techniques have been used to efficiently shuttle iron oxides into nonphagocytic cells, but all methods used until now require a prolonged incubation of cells. We hypothesized that instant magnetic labeling of cells could be achieved using electroporation. Neural stem cells (NSCs) and leukocytes from spleen and lymph nodes were suspended in a ferumoxide labeling solution, loaded into cuvettes, and subjected to electromechanical permeabilization using electroporation. Magnetically labeled cells were assayed for labeling efficiency, as well as for potential toxicity or altered function. To confirm the method's applicability to detect cells, MRI experiments were performed at 11.7 T. Magnetoelectroporation of NSCs, as demonstrated by Prussian blue staining, anti-dextran immunostaining, and a quantitative iron uptake assay, proved to be an efficient intracellular magnetic labeling method. Leukocytes including lymphocytes, which are notoriously difficult to label because of their membrane properties and small cytoplasmic volume, also demonstrated a pronounced uptake of ferumoxide. MRI experiments showed that labeled NSCs could be visualized as single cells and cell clusters in gelatin phantoms, and as proliferating cell masses in mouse brain. We have developed a convenient technique for instant magnetic labeling of cells. Because magnetoelectroporation allows the use of ferumoxides approved by the US Food and Drug Administration without additional agents, it has excellent potential for clinical translation.
authors.affiliation: Russell H. Morgan Department of Radiology and Radiological Science, Division of MR Research, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Piotr
authors.initials: P
authors.lastname: Walczak
authors.forename: Jesus
authors.initials: J
authors.lastname: Ruiz-Cabello
authors.forename: Dorota A
authors.initials: DA
authors.lastname: Kedziorek
authors.forename: Assaf A
authors.initials: AA
authors.lastname: Gilad
authors.forename: Sopo
authors.initials: S
authors.lastname: Lin
authors.forename: Bradley
authors.initials: B
authors.lastname: Barnett
authors.forename: Lu
authors.initials: L
authors.lastname: Qin
authors.forename: Hyam
authors.initials: H
authors.lastname: Levitsky
authors.forename: Jeff W M
authors.initials: JW
authors.lastname: Bulte
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D002470:Cell Survival; D018274:Electroporation / Q000379:methods*; D007962:Leukocytes / Q000166:cytology*; D008279:Magnetic Resonance Imaging / Q000379:methods*; D008280:Magnetics; D051379:Mice; D009474:Neurons / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*; D014481:United States; D014486:United States Food and Drug Administration / Q000331:legislation & jurisprudence*