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1.
Figure 7

Figure 7. From: Molecular Imaging of Bone Marrow Mononuclear Cell Homing and Engraftment in Ischemic Myocardium.

Ventricular function following BMMC therapy. (A) Representative M-mode images of hearts at 4 weeks following I/R injury with administration of PBS (left) versus BMMCs (right) (white scale bars = 5mm). (B) Quantification of left ventricular function demonstrates a trend towards improved functional recovery at 4 weeks post-I/R in animals receiving BMMCs compared to PBS (n=5–6 per time point) but did not achieve statistical significance.

Ahmad Y. Sheikh, et al. Stem Cells. ;25(10):2677-2684.
2.
Figure 1

Figure 1. From: Molecular Imaging of Bone Marrow Mononuclear Cell Homing and Engraftment in Ischemic Myocardium.

FACS analysis of bone marrow mononuclear cells (BMMCs) from L2G85 transgenic reporter mice reveals typical proportions of progenitor cells for the FVB strain. Green curves with corresponding percentages representing BMMC samples labeled with (A) CD31, (B) c-kit, (C) CD45, (D) Sca-1, and (E) CD34. Blue curves and percentages demonstrate negative controls (no antibody). (F) Approximately 4.5% of the BMMCs stain positive for both c-kit and Sca-1, similar to preparations used in clinical studies of BMMC transplantation in humans.

Ahmad Y. Sheikh, et al. Stem Cells. ;25(10):2677-2684.
3.
Figure 3

Figure 3. From: Molecular Imaging of Bone Marrow Mononuclear Cell Homing and Engraftment in Ischemic Myocardium.

Bioluminescence imaging demonstrates exogenously delivered BMMC preferentially home to injured myocardium. Images following the same animals (sham on left and I/R injury on right) for 4 weeks following intravenous delivery of L2G85-derived BMMCs (note the maximum values for scale bars in p/s/cm2/sr are different in the three rows). Persistently elevated signal from the area overlying the heart can be observed through day 14, followed by relatively similar decreasing trend in signal intensity by day 28. Images at day 10 demonstrate entrapment of cells in extra-cardiac sites such as the spleen (yellow arrows) and long bones of the lower extremities (red arrows).

Ahmad Y. Sheikh, et al. Stem Cells. ;25(10):2677-2684.
4.
Figure 5

Figure 5. From: Molecular Imaging of Bone Marrow Mononuclear Cell Homing and Engraftment in Ischemic Myocardium.

Histological evaluation confirms BMMC homing to the infarcted heart. Fluorescence microscopy images of a representative heart 2 days following 30 minutes of I/R injury and injection of 5×106 BMMN cells via tail vein. All panels stained with anti-troponin (red), anti-GFP (green), and DAPI (blue). (A) Infarcted area demonstrated by lack of bright troponin stain, with numerous GFP positive cells in the infarct border zone (scale bar = 50 μm). (B) High-power view demonstrating numerous GFP-expressing cells within the myocardium (scale bar = 10 μm). (C) Confocal laser microscopy image confirming presence of GFP-expressing cells within infarcted areas of the myocardium (scale bar = 5 μm).

Ahmad Y. Sheikh, et al. Stem Cells. ;25(10):2677-2684.
5.
Figure 4

Figure 4. From: Molecular Imaging of Bone Marrow Mononuclear Cell Homing and Engraftment in Ischemic Myocardium.

Ex vivo imaging confirms presence of transplanted cells within the myocardium. (A) Quantification of signals from ROIs over the thorax demonstrates significantly increased cell numbers in animals with I/R injury (white bars) compared to sham (gray bars) at days 2–6 following transplant (*P<0.05). Mean baseline Log10BLI (p/s/cm2/sr) of unoperated, uninjected mice was 3.3±2.1 (n=5 animals imaged at different time points). This was consistently 1–2 orders of magnitude lower than that of the highest signals attained in the animals receiving cells and I/R injury. (B) Ex vivo imaging of hearts two days following I/R injury with intravenous PBS (left), sham surgery with BMMCs (middle), or I/R injury with BMMCs (right) confirms homing of intravenously delivered BMMCs to the heart (white scale bars = 5 mm).

Ahmad Y. Sheikh, et al. Stem Cells. ;25(10):2677-2684.
6.
Figure 6

Figure 6. From: Molecular Imaging of Bone Marrow Mononuclear Cell Homing and Engraftment in Ischemic Myocardium.

Real time PCR quantification of surviving male transplanted cells within female hearts. (A) Plot of TaqMan-based Sry-quantification versus number of male cells in female hearts demonstrates a robust correlation between cycle count and known cell number (R2=0.99). (B) Ex vivo TaqMan analysis of hearts undergoing sham surgery (grey bars) versus I/R injury (white bars) demonstrates statistically significant increase of BMMCs homing in to injured hearts compared to sham at day 2 following transplant. This trend persists at week 2, but does not achieve statistical significance (n=5–6/group) (*P<0.05). Both results mirror findings generated by longitudinal bioluminescence imaging in vivo as shown in Figures 3 & 4.

Ahmad Y. Sheikh, et al. Stem Cells. ;25(10):2677-2684.
7.
Figure 2

Figure 2. From: Molecular Imaging of Bone Marrow Mononuclear Cell Homing and Engraftment in Ischemic Myocardium.

Reporter gene activity correlates robustly with cell number. (A) Bioluminescence imaging (BLI) of increasing numbers of BMMC in vitro (total cell count given above corresponding well with color scale bar representing range of signal in p/s/cm2/sr). (B) Correlation of cell numbers (x-axis) with BLI signal (left) and Fluc enzyme activity (right) demonstrate linear relationships with R2 values of 0.99. (C) Confocal laser microscopy of BMMCs demonstrates bright, cytosolic eGFP expression with corresponding nuclei stained blue with DAPI (scale bar = 5 μm). (D) FACS analysis of BMMCs from wild-type control FVB mouse (red) and L2G85 transgenic mouse (green) demonstrates robust eGFP expression by over 87% of the cells.

Ahmad Y. Sheikh, et al. Stem Cells. ;25(10):2677-2684.

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