Figure 1.
Clipping of M1 portion of left middle cerebral artery with small vascular clip.
(A) The M1 region (arrow) of the MCA was exposed by a retractor deep in the Sylvian fissure. Frontal is frontal lobe, Temporal is temporal lobe. (B) A small vascular clip was applied to the M1 portion. (C) MCA was occluded by a clip. (D) Before clipping, the color of venous blood was normal. (E) Darkning of cortical veins was observed during occlusion of MCA. (arrowheads). (F) After releasing the clip, the color of venous blood returned to normal.
Table 1.
Developing model summary (chronological order).
Table 2.
Developing model parameters.
Figure 2.
MRI of model development (initial) series.
Axial images of MRI FLAIR at day 3 obtained from all animals in the model development (initial) series. Upper panels (#1, #2, #4, #6) are collateral-poor animals, lower panels (#3, #5, #7, #11) are collateral-rich animals. High intensity area indicating cerebral infarction is primarily located in the left basal ganglia in collateral-rich animals. There was more extensive infarction areas in the MCA territory including cortex in collateral-poor animals.
Figure 3.
Scattered plots of stroke volume from model development series (n = 4/group).
Scattered plots show the stroke volume measured with axial images of MRI FLAIR at day 3. The stroke volume of collateral poor group (103.9±18.7 cm3) is greater than the collateral rich group (23.9±7.5 cm3; p = 0.0032). Each black symbol with number corresponds to the animal number in Table 1. Long symbols show averages. Given the small size, this was confirmed with a nonparametric test, the Mann-Whitney U test (p = 0.029).
Figure 4.
A representative animal of collateral poor NHP (2 hour occlusion of MCA).
Axial (A) and coronal (B) MRI FLAIR at day 3 demonstrate extensive high intensity areas located in the left MCA territory. Axial (C) and coronal (D) image of MRI T2WI at 1-year post-stroke demonstrated that high intensity areas remained. Gross brain anatomy revealed obvious lesion on the lateral surface of the brain 1 year after stroke (E, F).
Figure 5.
A representative animal of collateral rich NHP (2 hours occlusion of MCA).
Axial (A) and coronal (B) MRI FLAIR at day 3 demonstrate high intensity area located primarily in the left basal ganglia. Axial (C) and coronal (D) image of MRI T2WI at 1-year post- stroke demonstrated high intensity area is reduced. Gross brain anatomy revealed no obvious lesion in the surface of the brain at 1 year after stroke (E, F).
Table 3.
Safety Study animals summary.
Table 4.
Safety Study animals parameters.
Figure 6.
MRI FLAIR images of hMSC infused NHPs.
Axial images of available MRI of hMSC infused NHPs at day 3, day 7, 1 month, 6 months and 12 months. #12, #14, #17 and #18 correspond to the animal number in Table 3.
Figure 7.
MRI FLAIR images of serum infused NHPs.
Axial images of available MRI of serum infused NHPs at day 3, day 7, 1 month, 6 months and 12 months. #13, #15, #19, #20 and #22 correspond to the animal number in Table 3. #13 died at day 7.
Figure 8.
Summary of stroke volume from the safety study group.
Measured stroke volume (day 3, day 7, 1 month, 6 months and 12 months) in the hMSC and serum infused animals (A–E). Each small symbol with number represents a single animal. Long symbols are averages. Across all MRI methods, there was an overall decrease in stroke volume, but no statistical significant difference. Summarized time-course is shown in F.
Figure 9.
A representative hMSC infused NHP (#14).
(A) Coronal image of MRI FLAIR obtained at 12 months after infusion of hMSCs demonstrate high intensity area indicating stroke in the left basal ganglia and macroscopic image (B) shows necrotic cavity (arrow). Montage image (C) of HE staining of frozen coronal sections revealed cavity formation in the basal ganglia and no tumor formation. High power images of HE staining revealed infiltration of inflammatory cells in the cavity and fibrous gliosis in the wall of the cavity. No apparent infused hMSCs can be found in the lesion area. Scale bars = 1 mm (C), 200 µm (D–G).
Figure 10.
Neurological score in the hMSC and serum infused animals at day 1 (A), day 3 (B), day 7 (C), day 14 (D), 1 month (E), 6 months (F) and 12 months (G). Each small symbol with number represents a single animal. Long symbols indicate averages. There was an overall significant decrease in Neurological score in hMSC group, but no statistical significant difference between the groups at all time points. Summarized time-course is shown in H, I. Circled area in H is expanded in (I).