Distribution of hMSC to arteries/arterioles, veins and capillaries/end arterioles in the CAM.

<p><b>A.</b> Distribution of hMSC compared to lymphocytes and effects of pre-treatment with anti-SLeX and/or anti-α4 integrin (n = 5). <b>B</b>. Distribution in arteries of hMSC from 5 preparations from 5 different donors of marrow repeated 5 times.</p

3-Dimensional images of cells in rhodamine-labeled vesicles of chick embryo CAM.

<p>Orthologous projections of z-stacked photomicrographs of the CAM at 200× magnification. Crosshairs indicate cell of interest. <b>A.</b> B16F1 melanoma cells primarily embolized in the overlying capillary plexus (arrowhead) and at the ends of tapering arterioles. <b>B</b>. An hMSC, retaining its shape, adhered in a large vessel (dashed lines) lying beneath the capillary plexus.</p

Low passage hMSC express α4 integrin and SLeX.

<p>hMSC derived from four preparations from four different donors were assayed for expression of α4 integrin and SLeX by flow cytometry. Passage 1 cells were plated overnight to recover adherent viable cells and then re-plated at 100 cells/cm². The cells were harvested when 70 to 80% confluent.</p

Real time assay of cells in vessels of the chick embryo CAM.

<p><b>A.</b> Schematic for injecting cells or beads into a large vein of the CAM and capturing images for 3 to 10 minutes at either 40× or 100× magnification. <b>B. (upper panel).</b> Green B16F1 melanoma cells were primarily embolized in the capillary bed and had distorted morphology (∧). <b>(lower panel).</b> Green hMSC retained a regular morphology and were found both within arteries (†) and within the capillary beds (#). Images taken 10 minutes after injection of the cells. Arrows indicate direction of blood flow. Magnification 100×.</p

Clearance from the circulation of hMSC, melanoma cells and 10 µm inert beads.

<p>Inflexible inert 10 µM beads and B16F1 are cleared from circulation faster than hMSC. <b>A.</b> Values for cellular flux calculated as the average number of cells or 10 µm beads counted within vessels each minute in the CAM at 100× magnification. B. Values expressed as percentage flux were calculated as cells or beads in one minute as % of total observed in 10 minutes (n≥6).</p

Real-time PCR analysis of cytokine and chemokine profiles of EAEASCs and WtASCs.

<p>Both cell types were cultured overnight, trypsinized, and analyzed by real-time PCR to determine the expression levels of various cytokines and chemokines TNFα, IL-6, MCP-1, MIP-1α, RANTES, KC, MIP-2α and VEGF. # means <i>P</i><0.05 EAEASCs vs WtASCs (t-test, n = 3).</p

Clinical scoring for disease onset and progression.

<p>The clinical scores for the HBSS-treated (n = 20), EAEASC-treated (n = 20) and WtASC-treated (n = 20) EAE mouse groups were recorded daily for the duration of the study. WtASCs significantly reduced the clinical symptoms from PDI13 where the EAEASCs failed to mediate any therapeutic efficacy or improvement. # means <i>P</i><0.01 vs HBSS-treated EAE group (post-hoc, n = 20); • means <i>P</i><0.01 vs EAEASCs-treated EAE group (post-hoc, n = 20).</p

Lesion and cell infiltration analysis on the spinal cord.

<p>Spinal cords were collected at sacrifice from 3 mice per group. Each spinal cord was sectioned, mounted, and stained with anti-CD3, CD11b, and CD45 followed by Hematoxylin counterstain for lesion size, number and cell infiltration analysis. # means P<0.05 vs EAEASC-treated EAE group (post-hoc, n = 9); & means P<0.05 vs HBSS-treated EAE group (post-hoc, n = 9).</p

Pro-inflammatory cytokine protein levels in mouse serum.

<p>Sera from 5 mice per group at sacrifice were pooled and analyzed by ELISA to detect levels of TNF-α, IL-12, and IL-17. # means <i>P</i><0.05 vs HBSS-treated EAE group (post-hoc, n = 5); • means <i>P</i><0.05 vs EAEASCs-treated EAE group (post-hoc, n = 5); & means <i>P</i><0.05 vs normal naïve mouse group (post-hoc, n = 5).</p

Cell surface marker profiles of EAEASCs and WtASCs.

<p>A) Cells were analyzed by flow cytometry for MSC surface markers CD29, Sca1; hematopoietic markers CD34 and CD45; phagocytic lineage marker CD11b; and endothelial marker CD31. Gray filled: isotype control; gray line: WtASCs; black line: EAEASCs. B) Cell size based on forward scatter signal of flow cytometry (<i>P</i> = 0.26, t-test, n = 7).</p