Partec CyFlow MiniPOC.

<p>The Partec CyFlow MiniPOC counting device (A) and its display (B) in routine (left) or expert mode (right). Both images are used with permission of Partec/Sysmex GmbH, Germany.</p

Comparison of the CyFlow MiniPOC and CyFlow Counter.

<p>Bland-Altman plot comparing absolute CD4+ T cell counts obtained by Cyflow MiniPOC and CyFlow Counter by using the CD4 easy count kit (A) and the CD4% easy count kit (B).</p

Comparison of the CyFlow MiniPOC and CyFlow Counter.

<p>Linear Regression(A) and Bland-Altman Plot (B) for CD4+ T cell percent obtained by Cyflow MiniPOC and CyFlow Counter by using the CD4% easy count kit.</p

Comparison of the CyFlow MiniPOC and BC FC 500.

<p>Correlation and R² values for CD4+ T cell count (A) and percentage (B) obtained by Cyflow MiniPOC and BC FC 500 reference system.</p

Carry-over contamination analysis.

<p>The CD4 absolute count (A) and CD4% (B) were plotted as the mean value of three analysis sequences. Error bars indicate the standard deviation of three measurements.</p

Precision assessment.

<p>Bar charts with error bars indicating the standard deviation of CD4 T cell count (A) and CD4% (B) of a sample analyzed 10 times in single or in bulk. CV = Coefficient of variation.</p

hDPSCs surface antigens expression and <i>in vitro</i> multilineage differentiation ability.

<p>A: Cytofluorimetric analysis of Stro-1, c-Kit and CD34 expression in hDPSCs cultured in FCS-medium and in HS-medium. Dot plots reporting SSC vs fluorescence are shown. In the histograms, the net fluorescence value was calculated by linearizing the fluorescence value from the logarithmic scale and subtracting the linearized value of the unstained sample to the linearized value of the stained one. Data represent the mean±SD of three different experiments. * indicates values of paired <i>t</i>-test HS vs. FCS (*<i>p<</i>0.05). B: In the first line are shown double immunofluorescence images of hDPSCs/C₂C₁₂ co-culture stained by anti-hMit (green) and anti-myosin (red) Abs. DAPI staining is shown in blue. The second line shows oil red staining of hDPSCs differentiated for two weeks towards adipogenic lineage with HS or FCS supplemented medium. Cells were counterstained with Harris haematoxylin. Images, in third line represent anti-β3-Tubulin immunofluorescence labelling on hDPSCs differentiated in culture neurogenic media supplemented with FCS or HS. Bar 50 µm.</p

Confocal images of implants pre-differentiated in both the conditions.

<p>A–D: Double fluorescence signals from DAPI (blue) and anti-hMit Ab (green) superimposed to pseudo-phase contrast images (B, D). Arrowheads indicate cells entrapped in calcified bone matrix clearly stained by anti-human mitochondria antibody. E–H: triple fluorescence signals from DAPI (blue) and anti-hMit (green) and anti-OCN (red) Abs superimposed to pseudo-phase contrast images (F, H). Yellow arrowheads indicate osteocytes labelled by the two Abs; cyan arrowheads indicate OCN deposits in extracelluar bone matrix. I–L: triple fluorescence signals from DAPI (blue) and anti-hMit (green) and anti-von Willebrand factor (red) Abs superimposed to pseudo-phase contrast images (J, L). Arrowheads indicate vasa double stained by the two Abs. Bar: 10 µm.</p

Comparative histological analysis (haematoxylin/eosin staining) of the critical size cranial defect reconstruction by hDPSCs/collagen constructs 40 days post-surgery.

<p>Images show transversal sections carried out through the central area of the implants. A–C: cranial defect filled with hDPSCs/collagen pre-differentiated with FCS containing medium; D–F: cranial defect closed with hDPSCs/collagen pre-differentiated with HS containing medium; (dotted line delimitates the areas of bone resection; arrowheads indicate vasa; * indicate areas of active bone deposition). Bar: 100 µm. G: morphometric analysis of new-formed bone areas in controls (C), FCS and HS implants. Values are mean ± SD of the percentage of regenerated bone respect to the whole resected bone area. HS and FCS n = 6; C n = 4; white * inside the column indicate values of ANOVA test of HS and FCS vs. C (**<i>p<</i>0.01, ***<i>p<</i>0.001); black * indicate values of ANOVA test of HS vs. FCS (***<i>p<</i>0.001). H: number of vasa in the scaffold not yet reabsorbed and in new-formed bone areas. Data, normalized to areas of the scaffold not yet reabsorbed and of new-formed bone areas respectively, were presented as mean ± SD (vasa number respect to the total implant area) of each experimental group (controls n = 4; treated n = 6). White * inside the column indicate values of ANOVA test of HS and FCS vs. C (***<i>p<</i>0.001); black * indicate values of ANOVA test of HS vs. FCS (*<i>p<</i>0.05).</p

Kaplan-Meier survival curves related to the free-of-therapy period.

<p>The survival analysis was performed considering the time elapsed from PHI to the occurrence of failure, defined as the start of antiretroviral therapy. Upper pane, all patients; lower panels, patients with different degree of CD8+ T cell activation.</p