title: Internalisation of polymeric nanosensors in mesenchymal stem cells: analysis by flow cytometry and confocal microscopy.
abstract: The aim of this study was to demonstrate that flow cytometry and confocal microscopy could be applied in a complementary manner to analyse the internalisation of polymeric nanosensors in mesenchymal stem cells (MSC). The two techniques are able to provide en masse data analysis of nanosensors from large cell populations and detailed images of intracellular nanosensor localisation, respectively. The polyacrylamide nanosensors used in this investigation had been modified to contain free amine groups which were subsequently conjugated to Tat peptide, which acted as a delivery vector for nanosensor internalisation. Flow cytometry was used to confirm the health of MSC culture and assess the impact of nanosensor internalisation. MSC were characterised using fluorescently tagged CD cell surface markers that were also used to show that nanosensor internalisation did not negatively impact on MSC culture. Additionally it was shown that flow cytometry can be used to measure fluorophores located both on the cell surface and internalised within the cell. Complementary data was obtained using confocal microscopy to confirm nanosensor internalisation within MSC.
authors.affiliation: Laboratory of Biophysics and Surface Analysis, School of Pharmacy, University of Nottingham, University Park, Nottingham NG72RD, UK.
authors.forename: Paul G
authors.initials: PG
authors.lastname: Coupland
authors.forename: Karen A
authors.initials: KA
authors.lastname: Fisher
authors.forename: D Rhodri E
authors.initials: DR
authors.lastname: Jones
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Aylott
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D004337:Drug Carriers / Q000009:adverse effects / Q000737:chemistry*; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000009:adverse effects / Q000032:analysis*; D015696:Gene Products, tat / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000166:cytology / Q000196:embryology; D059630:Mesenchymal Stem Cells / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size
chemical_list: D003470:Culture Media; D004337:Drug Carriers; D005456:Fluorescent Dyes; D015696:Gene Products, tat