Characterization of EPCs, AD-MSCs and UC-MSCs.

<p>A, Expression patterns of endothelial makers on EPCs were analyzed by FACS. EPCs highly expressed CD31, vWF, CD144 and CD105, partly positive expressed VEGFR-2 and CD34, negative for CD90, CD45, CD14 and CD19. B, Biological function of EPCs was identified. a, Representative phase contrast images of cobblestone-like EPCs. b, EPCs bound with UEA-1 (red). c, EPCs incorporated DiI-Ac-LDL (green). d, EPCs formed vascular-like tubes on matrigel. C, Phenotype analysis of AD-MSCs and UC-MSCs by FACS. Both AD-MSCs and UC-MSCs were positive for CD29, CD90, CD73 and CD105, negative for VEGFR-2, CD14, CD31, CD34 and CD45. D, After 14 days of induction, AD-MSCs and UC-MSCs were differentiated into adipocytes and osteocytes. a, Adipogenic induction of AD-MSCs. b, Adipogenic induction of UC-MSCs. c, Osteogenic induction of AD-MSCs. d, Osteogenic induction of UC-MSCs. Adipogenesis was detected by the formation of neutral lipid vacuoles stainable with oil red O (red-orange). Osteogenesis was demonstrated by detection of alkaline phosphatase activity (brown).</p

AD-MSCs can promote vessel formation <i>in vivo</i>.

<p>After EPCs were implanted with/without UC-MSCs or AD-MSCs into SCID mice for 2 weeks, formed vessels in matrigel were subjected to immune-rejection by allo-PBMCs. The results were demonstrated by gross observation of the matrigel grafts (A) or by pathological section observation after H&E staining (B) or immunofluorescence staining (green: CD31; red: SMAα; blue: DAPI. 400×)(C). Implanted cells in matrigel formed vessels that inosculated with host vasculature and were perfused with mouse blood. EPCs implanted with AD-MSCs formed more vessels than EPCs implanted alone or EPCs implanted with UC-MSCs (D). The vessels in EPC+AD-MSC grafts also had significantly larger lumen than other groups (D). After allo-PBMC injection, red blood cell leakage was very obvious in EPC grafts or EPC+UC-MSC grafts (indicated by red arrows). In EPC+AD-MSC grafts, the vessels remained stable, and the red blood cell infiltration could hardly be observed.</p

Immuno-phenotyping of EPCs, AD-MSCs and UC-MSCs.

<p>A, Typical FACS results of MHC-I, MHC-II, CD40, CD80 and CD86 expression of three kinds of cells, with or without IFN-γ-treatment. B, Statistical analysis of FACS results of MHC-I, MHC-II and CD40 expression of the three kinds of cells, with or without IFN-γ-treatment. EPCs and AD-MSCs expressed significantly higher levels of MHC I and CD40 when compared with UC-MSCs. All three kinds of cells were negative for MHC II, CD80 and CD86. After stimulated with IFN-γ, the expression of MHC I, MHC II and CD40 were up-regulated in all the three kinds of cells, while the induced expression level in UC-MSCs was relatively low compared to AD-MSCs and EPCs (n = 3, P<0.01). C, mRNA levels of IFN-γR1 and IFN-γR2 gene were compared within three kinds of cells. The expression levels of both IFN-γR1 and IFN-γR2 were highest in EPCs, lowest in UC-MSCs (n = 3, P<0.01). D, Protein level of IFN-γR1 and IFN-γR2 were tested by Western blot, and compared within three kinds of cells with Quantity One software. The expression levels of both IFN-γR1 (EPC <i>vs</i> UC-MSC, P = 0.03) and IFN-γR2 (EPC <i>vs</i> UC-MSC, P = 0.016) were highest in EPCs, lowest in UC-MSCs (n = 3).</p

Primer sets of RT-PCR that used in this experiment.

<p>Primer sets of RT-PCR that used in this experiment.</p

The stimulating effect of EPCs, AD-MSCs and UC-MSCs on the proliferation of T cell subsets and the cytokine secretion of the three kinds of cells when co-cultured with allo-PBMCs.

<p>The stimulating effect of EPCs, AD-MSCs and UC-MSCs on the proliferation of T cell subsets was further analyzed. After one week of co-culture, CEDA-SE labeled T cells were stained with anti-CD4 or anti-CD8 antibody and subjected to FACS analysis. A and B, Typical results of 5 independent tests with similar trends. AD-MSCs, UC-MSCs or EPCs alone could hardly stimulate CD4+T or CD8+T proliferation, though the stimulating effect of EPCs was slightly higher. After pre-treatment by IFN-γ, EPCs could strongly stimulate CD4+T and CD8+T proliferation. However, this effect could be significantly inhibited by both AD-MSCs and UC-MSCs (n = 5, P<0.05). C, Cytokine secretion was analyzed after a 7-day co-culture of allo-PBMCs with different stimulators. EPCs can stimulate the production of IFN-γ and IL-10. AD-MSCs can down-regulate the secretion of IFN-γ (EPCs + IFN-γ group vs EPCs + IFN-γ + AD-MSCs group) and up-regulate the secretion of IL-10 (blank vs ADMSCs group, EPCs group vs EPCs + AD-MSCs group) (n = 3, P<0.05).</p

AD-MSCs and UC-MSCs could inhibit the proliferation of allo-PBMCs <i>in vitro</i>.

<p>Proliferation of allo-PBMCs co-cultured with different stimulating cells was analyzed by CFDA-SE-based proliferation assay. A, Typical results of the mixed lymphocyte reaction (MLR) assay. a-l show allo-PBMC proliferation after co-culture with different stimulators: a, blank as negative control; b, AD-MSCs; c, UC-MSCs; d, PHA as positive control; e, AD-MSCs + PHA; f: UC-MSCs + PHA; g, EPCs; h, EPCs + AD-MSCs; i, EPCs + UC-MSCs; j, IFN-γ-treated EPCs; k, IFN-γ-treated EPCs + AD-MSCs; l, IFN-γ-treated EPCs + UC-MSCs. B and C, The statistical results of MLR assay. B, AD-MSCs and UC-MSCs could slightly stimulate allo-PBMC proliferation. EPCs displayed a significantly stronger stimulating effect than AD-MSCs and UC-MSCs (n = 5, P<0.05). C, IFN-γ-treated EPCs showed a higher stimulating effect than EPCs. AD-MSCs and UC-MSCs could strongly down-regulate the stimulating effect of EPC and IFN-γ-treated EPCs. UC-MSCs have a more remarkable immune-inhibiting effect on IFN-γ-treated EPCs when compared with AD-MSCs (n = 5, P<0.05).</p

Ordered Porous Pd Octahedra Covered with Monolayer Ru Atoms

Monolayer Ru atoms covered highly ordered porous Pd octahedra have been synthesized via the underpotential deposition and thermodynamic control. Shape evolution from concave nanocube to octahedron with six hollow cavities was observed. Using aberration-corrected high-resolution transmission electron microscopy and X-ray photoelectron spectroscopy, we provide quantitative evidence to prove that only a monolayer of Ru atoms was deposited on the surface of porous Pd octahedra. The as-prepared monolayer Ru atoms covered Pd nanostructures exhibited excellent catalytic property in terms of semihydrogenation of alkynes