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3562 Chung et al.

Fig. 11. In vivo detection of CM-DiI-labeled HUCB-derived Fig. 13. Polymorphologic human cell nuclei marker-expressing cells.
MSCs. CM-DiI labeled HUCB-derived MSCs are shown as red Human cell nuclei marker (MAB1281)-expressing cells appear as
fluorescent cells and are observed at 4 weeks after transplantation in brown cells (arrows) and are aggregated around lesion area. These
penumbra region (within rectangle). 3200. [Color figure can be cells show polymorphologic features. Scale bar 5 100 lm. [Color
viewed in the online issue, which is available at www.interscience. figure can be viewed in the online issue, which is available at
wiley.com.] www.interscience.wiley.com.]

marker (MAB1281) was used to confirm that trans-
planted HUCB-derived MSCs had migrated to the
infarction lesion area. Human cell nuclei marker-express-
ing cells were observed around the infarction lesion area
in the penumbra region by fluorescent microscopy
(Fig. 12). Human cell nuclei marker-expressing cells
were observed mainly in the left hemisphere section
with the brain infarction lesion. Human cell nuclei
marker (MAB1281)-expressing cells were examined
using immunohistochemical analysis and a counterstain
procedure. Human cell nuclei marker-expressing cells
were aggregated around the lesion area and had
polymorphologic features (Fig. 13).

Immunofluorescence and
Immunohistochemical Studies
Immunofluorescence and immunohistochemical
Fig. 12. Fluorescence microscopy images of HUCB-derived MSCs of studies demonstrated that the CM-DiI-positive donor
left ischemic hemisphere (A) and right intact hemisphere (B). A-1,B-1: HUCB-derived MSCs expressed NeuN, GFAP, and
3100, A-2,B-2: 3400. A-1 and A-2 images are obtained from the left vWF. CM-DiI-positive cells and human cell nuclei
hemisphere with infarction lesion. Blue-fluorescent human nuclei marker (MAB1281)-expressing cells were also immuno-
marker-expressing cells are observed in the penumbra region (A-1: reactive for NeuN and GFAP (Figs. 14, 15). CM-DiI-
box, A-2: arrows). B-1 and B-2 images are obtained from the right positive cells were found in and around endothelial cells
intact hemisphere. No blue fluorescent cells are observed. In B-2, that were positive for vWF (Fig. 16). Confocal images
canine neuronal cells are not reactive with human nuclei antibody, showed expression of BDNF and VEGF in transplanted
so vacuoles are observed where neuronal cells are located. [Color HUCB-derived MSCs (Fig. 17).
figure can be viewed in the online issue, which is available at www.
interscience.wiley.com.]
DISCUSSION
A canine cerebral ischemic model was used because
In Vivo Detection of HUCB-Derived MSCs the intravascular approach to the internal carotid artery
Intraarterially administered, CM-DiI-prelabeled, by an endovascular interventional approach could be
HUCB-derived MSCs had migrated to and accumulated reasonably accomplished. Occlusion of the MCA and
around ischemic lesions in the penumbra area 4 weeks induction of cerebral ischemia in experimental dogs
after transplantation (Fig. 11). A human cell nuclei prior to the transplantation procedure were successful.

Journal of Neuroscience Research