Fig. 5
In vivo differentiation potential of Ptf1a-reprogrammed miNSCs. a miNSC10 cells were infected with lentiviruses containing CMV promoter-driven GFP and neomycin. After screening with G418, most miNSC cells expressed GFP as shown. b GFP-tagged miNSC10 cells were injected into the hippocampus region of 2-month-old mice. One month after transplantation, GFP+ cells were found successfully integrated into the tissue. Some of them were immunoreactive for the oligodendrocyte cell marker Olig2. c–f Transplanted GFP + cells immunoreactive for neuronal cell markers Dcx, NeuN, GABA, or vGLUT3. g Transplanted GFP+ cells co-expressing the astrocyte cell marker GFAP. h Immunolabeling for the presynaptic protein marker synapsin. i Single confocal plane images of the region outlined by the small rectangle in h show direct contact between the GFP + dendrite and synapsin + presynaptic terminals of the surrounding endogenous cells. j The 3D surface of confocal z-stacks of the region outlined by the large rectangle in h shows the formation of synaptic connections with endogenous cells. k The 3D surface of z-stacks of a dendrite shows the development of dendritic spines with typical head–neck structures (indicated by asterisks). Cells in b–h were counterstained with nuclear DAPI. Large arrows in b–g point to representative colocalized cells and/or processes. Small arrows in i and j indicate direct contact between neurites and multiple synapsin + presynaptic teminals. l Quantification of GFP + cells that are immunoreactive for NeuN, GABA, vGLUT3, GFAP, or Olig2. Scale bars, 80 μm (a) and 20 μm (b–h)
