Results of ELISA analysis for VEGF and GDNF.

<p>(A, B) In the lesioned striatum, VEGF was significantly increased by SCS at 1 week after 6-OHDA lesion (*p<0.05). At 2 weeks after 6-OHDA lesion, VEGF level in the lesioned striatum also appeared elevated, but did not reach statistical significance. (C, D) GDNF in the striatum of both sides was not significantly increased by SCS at 1 and 2 weeks after 6-OHDA lesion. (C-lesion: control group lesioned side Striatum; C-intact: control group intact side Striatum; S-lesion: 50 Hz SCS group lesioned side Striatum; S-intact: 50 Hz SCS group intact side Striatum, n = 10, respectively).</p

TH immunostaining in the substantia nigra pars compacta (SNc), and the ratio to the intact side.

<p>(A) TH immunostaining in the intact SNc. Severe loss of TH-positive neurons was seen in the lesioned side SNc of control group. Preservation of TH-positive neurons was seen at the lesioned side SNc of 50 Hz SCS group. Scale bar: 200 µm. (B) Significant preservation of TH-positive neurons in the lesioned-side SNc of 50 Hz SCS group, compared to those of control group (*p<0.05, n = 10, respectively).</p

Time course and SCS electrode, and the brain region punched out for protein assay.

<p>(A) Scheme showing overall experimental design. (B) Scheme showing experimental design for protein assay. (C) Photograph showing SCS electrode used in this study (diameter: 2 mm; wire length: 60 mm). (D) Scheme showing a rat during stimulation. (E) Brain tissue (diameter: 3 mm showing gray circle), corresponding to the striatum, was punched out from both the lesioned and the intact side.</p

Tyrosine hydroxylase (TH) immunostaining in the striatum and the ratio to the intact side.

<p>(A) TH immunostaining in the striatum. Severe loss of TH-positive fibers was seen in the lesioned striatum of control group. Preservation of TH-positive fibers was seen in the lesioned striatum of all SCS groups. Scale bar: 200 µm. (B) The all SCS groups showed significant preservation of TH-positive fibers in the lesioned striatum, compared to those in control group (*p<0.05, n = 10, respectively).</p

The results of cylinder test and amphetamine-induced rotation test.

<p>(A) Rats receiving 2 Hz and 50 Hz SCS showed reduction of the contralateral bias at 2 weeks after 6-OHDA lesion, compared to that of rats in control group. (B) The number of amphetamine-induced rotations in all SCS groups decreased, compared to that of control group. There was a significant amelioration in 50 Hz SCS group, compared to control group (*p<0.05, n = 10, respectively).</p

Reduction of infarct volumes after transplantation.

<p>A, B: Photographs of coronal brain sections of rats in control group (A) and 24h group (B) with 2,3,5-triphenyltetrazolium chloride (TTC) staining. C: Bar graph showing infarct volumes in each group. The infarct volumes of rats in 24h group significantly decreased compared to rats in control, 1h, and 6h groups at 7 days after MCAO (* p<0.05, n = 6 in control group, 5 in 1h group, 6 in 6h group, 6 in 24h group, 6 in 48h group, respectively). The infarct area ratio of rats in 48h group tended to decrease compared to control group, 1h group, and 6h group, but there was no statistical difference.</p

Migration of integrated MSCs in vivo.

<p>A, B: Photographs showing integrated MSCs in the brain. Integrated MSCs were mainly detected in the ischemic penumbra at 7 days after MCAO. Scale bar: 20 μm. C: Bar graph showing the number of integrated MSCs in the brain of rats in each group. The number of MSCs was significantly higher in 24h group than other transplantation groups (* p value<0.05, n = 10 in 1h group, 5 in 6h group, 9 in 24h group, 8 in 48h group, respectively).</p

Results of ELISA analyses for bFGF and SDF-1α.

<p>A: The bFGF level increased in the infarcted cortex of rats in 24h group. There was significant difference of the bFGF level between the infarcted cortex and the intact cortex of rats in 24h group and the infarcted and intact cortex of rats in control group. B: In the infarcted striatum, bFGF level did not increase by the transplantation (C-infarct: the infarct side of rats in control group; C-intact: the intact side of rats in control group; 24h-infarct: the infarct side of rats in 24h group; 24h-intact: the intact side of rats in 24h group, *p value<0.05, n = 8, respectively). C: SDF-1α level significantly increased in the infarcted cortex of rats in 24h group, compared to all other regions. D: In the infarcted striatum, SDF-1α level did not increase by the transplantation (* p value<0.05, n = 8, respectively).</p

Detection of labeled MSCs in vitro.

<p>A, B: Photographs of labeled MSCs <i>in vitro</i>. Fluorescent microscope detected MSCs with red-colored cytoplasm. Scale bar: A: 40μm; B: 20μm. The cell labeling rate was 71.4±5.7% at 2 hours after cell labeling (n = 12).</p

Intra-arterial MSC transplantation ameliorates stroke-induced neurological deficits.

<p>Neurological performance as revealed by mNSS shows that intra-arterial MSCs at 24h and 48h transplantation groups was better than that of control group. Rats in 24h group showed significant recovery, compared to all other groups at 3 and 7 days after MCAO (* p value<0.05 vs. control; ** p value<0.05 vs. all other groups, n = 17 in control group, 10 in 1h group, 11 in 6h group, 12 in 24h group, 12 in 48h group, respectively).</p