title: Intracellular labeling of mouse embryonic stem cell-derived neural progenitor aggregates with micron-sized particles of iron oxide.
abstract: OBJECTIVE
  Pluripotent stem cell (PSC)-derived neural progenitor cells (NPCs) represent an unlimited source for the treatment of various neurological disorders. NPCs are usually derived from PSCs through the formation of embryoid body (EB), an aggregate structure mimicking embryonic development. This study investigated the effect of labeling multicellular EB-NPC aggregates with micron-sized particles of iron oxide (MPIO) for cell tracking using magnetic resonance imaging (MRI).
  
  
  METHODS
  Intact and dissociated EB-NPC aggregates were labeled with various concentrations of MPIOs (0, 2.5, 5 and 10 μg Fe/mL). The labeled cells were analyzed by fluorescent imaging, flow cytometry and in vitro MRI for labeling efficiency and detectability. Moreover, the biological effects of intracellular MPIO on cell viability, cytotoxicity, proliferation and neural differentiation were evaluated.
  
  
  RESULTS
  Intact EB-NPC aggregates showed higher cell proliferation and viability compared with the dissociated cells. Despite diffusion limitation at low MPIO concentration, higher concentration of MPIO (i.e., 10 μg Fe/mL) was able to label EB-NPC aggregates at similar efficiency to the single cells. In vitro MRI showed concentration-dependent MPIO detection in EB-NPCs over 2.0-2.6 population doublings. More important, MPIO incorporation did not affect the proliferation and neural differentiation of EB-NPCs.
  
  
  CONCLUSIONS
  Multicellular EB-NPC aggregates can be efficiently labeled and tracked with MPIO while maintaining cell proliferation, phenotype and neural differentiation potential. This study demonstrated the feasibility of labeling EB-NPC aggregates with MPIO for cellular monitoring of in vitro cultures and in vivo transplantation.
authors.affiliation: Department of Chemical and Biomedical Engineering FAMU-FSU College of Engineering, Florida State University, Tallahassee, Florida, USA.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Sart
authors.affiliation: The National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Fabian Calixto
authors.initials: FC
authors.lastname: Bejarano
authors.affiliation: The National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Baird
authors.affiliation: Department of Chemical and Biomedical Engineering FAMU-FSU College of Engineering, Florida State University, Tallahassee, Florida, USA.
authors.forename: Yuanwei
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Department of Chemical and Biomedical Engineering FAMU-FSU College of Engineering, Florida State University, Tallahassee, Florida, USA; The National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
authors.forename: Jens T
authors.initials: JT
authors.lastname: Rosenberg
authors.affiliation: Department of Chemical and Biomedical Engineering FAMU-FSU College of Engineering, Florida State University, Tallahassee, Florida, USA.
authors.forename: Teng
authors.initials: T
authors.lastname: Ma
authors.affiliation: Department of Chemical and Biomedical Engineering FAMU-FSU College of Engineering, Florida State University, Tallahassee, Florida, USA; The National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA. Electronic address: scgrant@fsu.edu.
authors.forename: Samuel C
authors.initials: SC
authors.lastname: Grant
authors.affiliation: Department of Chemical and Biomedical Engineering FAMU-FSU College of Engineering, Florida State University, Tallahassee, Florida, USA. Electronic address: yli@eng.fsu.edu.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D058948:Cell Tracking / Q000379:methods; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D005290:Ferric Compounds / Q000494:pharmacology*; D008279:Magnetic Resonance Imaging / Q000379:methods; D058185:Magnetite Nanoparticles / Q000008:administration & dosage*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects; D010316:Particle Size; D013194:Staining and Labeling / Q000379:methods
chemical_list: D005290:Ferric Compounds; D058185:Magnetite Nanoparticles; C000499:ferric oxide
keywords: cell tracking; iron oxide particles; neural progenitor aggregates; pluripotent stem cells