Fig. 2
From: Development of a novel imaging system for cell therapy in the brain
Generation of Hipp-NSCs expressing the hNIS. a Map of the HIV-based lentivector CD11B-1 (System Biosciences). The hNIS cDNA was cloned in the multiple cloning site (MSC) located downstream of the CMV promoter. Downstream of the expression cassette for the hNIS, an EF1 promoter drives the expression of GFP. b Expression of GFP in Hipp-NSCs transduced with pseudoviral particles for 48 h. The fluorescent image is superimposed on the phase-contrast image. c Flow cytometric analysis of Hipp-NSCs transduced with the hNIS showing the expression of GFP (indicative of the expression of hNIS) in 62 % of the cells before sorting and in 96 % of the cells after sorting (d). e Immunofluorescence analysis of the expression of the hNIS (in red) in Hipp-NSCs transduced with the lentivector and selected by sorting for GFP. Nuclei are stained blue with DAPI. f Expression of GFP in differentiated Hipp-NSCs transduced with the hNIS. g Immunoreactivity for the hNIS in differentiated NIS-Hipp-NSCs. Nuclei are counterstained blue with DAPI. h Uptake of 99mTc in NIS-Hipp-NSCs was measured as counts per minute (CPM) in a gamma counter and corrected by subtracting background CPM. N = 3; *P < 0.01 by Student’s t test. Scale bars = 100 μm. CMV cytomegalovirus, DAPI 4′,6-diamidino-2-phenylindole, GFP green fluorescent protein, Hipp-NSC hippocampus-derived neural stem cell, hNIS human sodium iodide symporter, NIS-Hipp-NSC hippocampus-derived neural stem cell expressing the human sodium iodide symporter, 99m Tc technetium-99m