Newly generated hNSCs.

<p>(A) Schematic representation of the designed vector system. The two imaging reporters Luciferase 2 (Luc2) and green fluorescence protein (GFP) are kept under the control of the constitutive active promoter EF1α and are linked via the T2A peptide sequence to ensure equal expression level of the two proteins. (B) Representative microscopic image of transduced and FACS sorted hNSCs. The overlay of the bright-field and fluorescence image is shown right. Scale bar: 50 μm</p

Longitudinal evaluation of cell viability by <i>in vivo</i> BLI.

<p>(A) BL images of unlabeled (left) and labeled (right) hNSCs, implanted in the right striatum and longitudinally evaluated from day 0 to day 9 post implantation [¹⁹F labeled cells day 0 (n = 9), day 1 (n = 9), day 2 (n = 8), day 5 (n = 8), day 7 (n = 7), day 9 (n = 5) / unlabeled cells day 0–9 (n = 4)]. (B) SBR (signal to background ratio) normalized to the first time point shows a decrease of cell viability within one week. (+) outliers at least 3x interquartile range.</p

<i>In vivo</i>¹⁹F MRI.

<p>(A) High resolution ¹H MR image (left) was acquired as anatomical reference and ¹⁹F MR image (center) was then acquired to localize the implanted cell graft. ¹H and ¹⁹F images were superimposed to combine anatomical information and spatial graft localization (right). (B) Two animals with quantitative depiction of 19F-labelled detectable cells at both two and eight days post implantation. C) Quantification of hNSCs labelled with PFPE at both, day 2 and day 8.</p

Histological analysis of graft survival and immune response of host tissue.

<p>An overview of the mouse brain slice (scale bar: 400 μm) and higher magnification confirm that Iba1 positive cells surround the cell graft (4x magnification, scale bar: 200 μm / 10x magnification, scale bar: 50 μm / 60x magnification, scale bar: 10 μm). GFP-transgene expression (green) and immunostainings with antibodies against: Iba1 (IBA), immunoreaction and HuNu, human nuclei marker. In the lower row 3D images of the IBA staining illustrate the surrounding of the cell graft by the immune cells.</p

Immunohistochemistry validation of grafted hNSCS.

<p>Histology of transplanted H9-EF1-Luc2-GFP cells either labeled with ¹⁹F (n = 4) (A) or unlabeled (n = 4) (B) 9 days after transplantation. An overview of the mouse brain slice (scale bar: 400 μm) and higher magnification of the grafted cells verified the localization of the transplanted cells (4x magnification, scale bar: 200 μm / 10x magnification, scale bar: 50 μm / 60x magnification, scale bar: 10 μm). GFP-transgene expression (green) and immunostainings with antibodies against: DCX, neuronal marker; HuNu, human nuclei marker; Mito, human mitochondria; GFAP, astrocyte marker; Luc, luciferase marker.</p

<i>In vitro</i> detectability of ¹⁹F labeled hNSCs by means of ¹⁹F MRI and ¹⁹F MRS.

<p>(A) ¹⁹F MRS of labeled hNSCs and a KF solution as internal standard to quantify the amount of ¹⁹F atoms per cell (B) high resolution ¹H MR image (left), acquired during the same session of the labeled cells ¹⁹F MR image (center). ¹H and ¹⁹F images are then merged to obtain a correct spatial localization (right).</p

Effect of the transduction and ¹⁹F labeling on hNSCs.

<p>(A) Cell viability is shown for WT hNSCs and transgenic EF1-Luc2-GFP hNSCs with and without ¹⁹F labeling (n = 6–8). (B) Cell proliferation was compared among different cell lines. The values were normalized to the WT hNSCs and expressed in percentage (n = 5). (C) <i>In vitro</i> BLI signal from transgenic unlabeled hNSCs compared to ¹⁹F labeled cells. (D) <i>In vitro</i> BLI signal is displayed for a dilution series of cells (labeled and unlabeled) in 6 independent experiments. (+) outliers at least 1.5x interquartile range.</p

KPF₆ 14.2 mM perfusion induced large amplitude depolarization events that propagated through the cortical mantel, from the hippocampus to the entorhinal (EC) and piriform cortex (PC).

<p>These events were interpreted as spreading depolarization phenomena (Somjen 2001). </p

A: Seizure occurrence after perfusion of the <i>in</i><i>vitro</i> isolated guinea pig brain with different K⁺ salt solutions.

<p>The white columns represent the total number of experiments. The light grey, dark grey and black columns mark the effects of the perfusion of K⁺ salts at 8, 14.2m 20 mM, respectively. <b>B</b>: Time at onset (black columns) and duration (white columns) of seizure activity after perfusion with 14.2 KPF₆, 14.2 mM KClO₄ and 20 mM KBF₄. <b>C</b>: Brain parenchyma concentration of two salts (KPF₆ and KBF₄) estimated by ¹⁹F MR spectroscopy on CA1-EC specimens, collected after arterial perfusion with 14.2 mM KPF₆ (n= 4; black column) and 20 mM KBF₄ (n= 3; grey column). </p

Effects of 4.2 mM KPF₆ (A), 14.2 mM KClO₄ (B) and 20 mM KCl (C) on the field potentials evoked in the piriform cortex by lateral olfactory tract (LOT) stimulation.

<p>Left, middle and right traces: before, during and after washout of the K⁺ salts, respectively.</p