<i>In vivo</i> images and immunohistochemical analysis of third and fourth generation MIN-O Line D/EmGFP lesions.

<p>a & d) Spectrally unmixed image overlaid on the corresponding monochrome image showing the presence of a GFP positive lesions in the general region of the 4^th right mammary fat pad of third (a) and fourth (d) generation MIN-O Line D/EmGFP. Images were acquired 33 and 29 days post-transplant, respectively. b) Immunohistochemical analysis of third generation lesion reveals weakened general GFP expression as well as several GFP-negative regions. e) Similar analysis of fourth generation lesion reveals large regions of GFP-negative cells. c & f) Enlarged sections of 5b and 5f, respectively. Arrows show EmGFP positive basal cells and arrowheads show EmGFP positive luminal cells.</p

<i>In vivo</i> image and immunohistochemical analysis of second generation MIN-O Line D/EmGFP lesion.

<p>a) Spectrally unmixed image overlaid on the corresponding monochrome image showing the presence of an EmGFP positive lesion in the general region of the 4^th right mammary fat pad. Image was acquired 29 days post-transplant. b) Immunohistochemical analysis of second generation lesion reveals strong general EmGFP expression, but also EmGFP-negative regions. c) Enlarged section of 4b showing a high degree of segregation of EmGFP-positive (upper right) and EmGFP-negative (lower left) cells. d) Enlarged section of 4b showing interspersed EmGFP-positive and EmGFP-negative cells. Arrows show EmGFP positive basal cells and arrowheads show EmGFP positive luminal cells.</p

<i>In vivo</i> and <i>ex vivo</i> images of first generation MIN-O Line D/EmGFP subline.

<p>a) Spectrally unmixed image overlaid on the corresponding monochrome image showing the presence of an EmGFP positive lesion in the general region of the 4^th right mammary fat pad. b) <i>Ex vivo</i> validation of <i>in vivo</i> image confirming the source of the EmGFP signal to be the lesion excised from the 4^th right mammary fat pad (right). For comparison, an EmGFP-negative lesion is shown on the left. Both images were acquired 27 days post-transplant.</p

Immunohistochemical analysis of excised MIN-O Line D/EmGFP lesion.

<p>Four micron sections of first generation MIN-O/EmGFP lesion either stained with Mayer’s H & E (a&b) or probed with α-GFP in order to asses distribution of EmGFP within the lesion (c&d). Evident in the IHC are morphological characteristics common to the parental MIN-O transplant line which include growth into the fat pad stroma, basal and luminal differentiation, remodeled stroma and increased vascularity (arrow). Analysis of EmGFP expression reveals a high degree of homogeneity throughout the MIN-O tissue (arrow), but not in the host-derived stroma (arrowhead).</p

Quantitative Real-Time PCR Analysis of Genomic EmGFP and PyVmT Sequence in Laser Captured EmGFP Positive and Negative Regions.

<p>a-d) Images acquired during laser capture of MIN-O tissue shows EmGFP negative regions before (a) and after (b) dissection and EmGFP positive regions before (c) and after (d) dissection. e) qPCR analysis of EmGFP and PyVmT sequence in genomic DNA isolated from laser captured sections. Calculated PFAFFL values were averaged for each primer pair and normalized (EmGFP positive tissue  = 100), allowing for direct comparison of the percent relative abundance of EmGFP and PyVmT sequence in designated samples. While genomic DNA from EmGFP negative tissue shows significant loss of EmGFP coding sequence, a similar decrease in sequence encoding PyVmT was not observed. These data indicate that, while the majority of tissue in both regions is MIN-O in origin, the EmGFP expression cassette has been largely expelled from the genome of EmGFP negative cells.</p

Short-term tracking of hPBSC radiolabeled with ⁸⁹Cu-PTSM by PET.

<p>hPBSC were radiolabeled with 20 µCi/mL of ⁶⁴Cu-PTSM. Cells radiolabeled with ⁶⁴Cu-PTSM were detected in the lung (white arrow) and liver (red arrow) on the day of postnatal transplant (day 0). Cells were observed in the liver (red arrow) and spinal column (yellow arrow) 24 h post-injection (day 1).</p

BLI and PET imaging of transplanted hPBSC.

<p>hPBSC expressing firefly luciferase were transplanted prenatally in the late first trimester <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077148#pone.0077148-Tarantal2" target="_blank">[7]</a>. At ∼3 months postnatal age D-luciferin was injected intravenously and BLI was performed to confirm the anatomical location of transplanted cells, then ⁸⁹Zr-Df-CD45 was injected. Monkey #1 showed a high level of bioluminescence in the abdominal region. Both animals showed strong PET signals on day 5 post-injection of ⁸⁹Zr-Df-CD45 within the liver. The muscular component of the peritoneum (white and yellow arrows) of Monkey #1 showed corresponding BLI and PET signals.</p

⁶⁴Cu-TETA-CD45 radiolabeling of hPBSC.

<p>hPBSC were radiolabeled with 0, 20, 40, 80, or 160 µCi/mL of ⁶⁴Cu-TETA-CD45. No significant changes in cell viability or degree of labeling were observed with increasing concentrations. A decline in cell growth and colony formation was observed when cells were incubated with ⁶⁴Cu-TETA-CD45 at a concentration >20 µCi/mL.</p

Anti-oxLDL Individual Serum Concentration-Time Profiles Measured by Assay B.

<p>Anti-oxLDL Individual Serum Concentration-Time Profiles Measured by Assay B.</p

PET Images from One Representative Animal at 1 hour Post Dose: Frontal View (A) and Sagittal View (B).

<p>PET Images from One Representative Animal at 1 hour Post Dose: Frontal View (A) and Sagittal View (B).</p