Journal: Cellular and molecular gastroenterology and hepatology

Article Title: Additive Effect of CD73 Inhibitor in Colorectal Cancer Treatment With CDK4/6 Inhibitor Through Regulation of PD-L1.

doi: 10.1016/j.jcmgh.2022.07.005

Figure Lengend Snippet: Figure 8. Negative corre- lation between ecto- enzymes and CCND1 expression in CRC. (A) IHC staining of human normal colon (n ¼ 40) and colon adenocarcinoma (n ¼ 64) specimens in a tissue microarray for CCND1 and CD163 (a macrophage marker). (B) Relative intensities of CCND1 staining in the lamina propria of a normal and cancerous colon. **P < .01 (unpaired t test). (C) Correlation analysis of NT5E and CCND1 expres- sion in the TCGA colorectal adenocarcinoma dataset. ***P < .001 (Student t test).

Article Snippet: For the fluorescence-activated cell sorting analysis, cells were stained with APC,-anti human CD163 (clone: GHI/61), PD-anti human CTLA4 (clone: L3D10), Alexa 647-anti human IDO1 (clone: 2E2/IDO1), PE-anti human DR4 (clone: DJR1), FITC-anti human CD47 (clone: REA220, FITC-anti human MICA&B (clone: 6D4), PE-anti human PD-L1 (clone: MIH2), PD-anti human CD69 (clone: FNM50), FITCanti human CD2 (clone: RPA-2.10), FITC-anti human CD20 (clone: 2H7), APC-anti mouse F4/80 (clone: BM8), Brilliant Violet 421TM-anti mouse CD11b (clone: M1/70), Brilliant Violet 570TM-anti mouse CD45 (clone: 104), PE-anti-human CD45 (clone: HI30), monoclonal antibodies (Biolegend; San Diego, CA, USA), FITC-anti-human CD40 (clone: REA733), PE-anti human CD80 (clone: 2D10), PE/vio770-anti human CD206 (clone: DCN228), PE-anti human CD62E (clone: REA280), PE-anti human CD192 (clone: REA264), APC-anti human I-CAM (clone: REA266), APC-anti human HLA-DR, DP, DQ (clone: REA332), APC-anti human CCL2 (clone: REA485), APC-anti human CD14 (clone: HI30), Vioblue-anti human CD31 (clone: TUK4), FTIC-anti-human CD86 (clone: FM95), PE-anti human CD73 (clone: AD2), PE-anti mouse PD-L1 (clone: 60533), PE-anti mouse CD39, and PE-anti human CD39 (clone: REA739) monoclonal antibodies (Miltenyi Biotec; Bergisch Gladbach, Germany), and PE/CF594anti human CD3 (clone: UCHT1), PE-anti human CD44 (clone: G44-26), PE-anti human CD183 (clone: 150503), and PD-1 (clone: REA739) monoclonal antibodies (BD Biosciences; San Jose, CA, USA) following the manufacturer’s protocol.

Techniques: Expressing, Immunohistochemistry, Microarray, Marker, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Human placenta-derived mesenchymal stem cells stimulate neuronal regeneration by promoting axon growth and restoring neuronal activity

doi: 10.3389/fcell.2023.1328261

Figure Lengend Snippet: Characterization of hPMSCs. (A) Morphology at passage 3. Both normoxic and hypoxic cultures show highly homogeneous spindle-like shape. Scale bar: 500 µm (B) Proliferation capacity. Cumulative population doublings of hPMSCs grown under hypoxia (square) are higher than cultures grown in normoxia (dot). (C) Flow cytometry shows specific hMSCs marker expression pattern: positive for CD105, CD44, CD90, CD73 and negative for CD14, CD19, CD34, CD45 and HLA-DR. (D) Osteogenic differentiation was confirmed by Alizarin red staining of calcium deposits. (E) Adipogenesis differentiation was followed by Oil Red O staining of lipids vacuoles (black arrows indicate lipid droplets). (F) Alcian Blue staining of proteoglycans demonstrated chondrogenesis differentiation. Scale bar: 100 μm. Two-way ANOVA and post hoc Sidak test for multiple comparisons between means (** p ≤ 0.005; * p ≤ 0.05).

Article Snippet: Antibodies against the following human antigens were used: CD105-FITC (Miltenyi Biotect, Bergisch Gladbach, Germany, cat# 130-112-327, 1:50), CD90-FITC (Miltenyi Biotec, cat# 130-114-901, 1:50), CD44-VioBlue (Miltenyi Biotec, cat# 130-113-906, 1:50), CD73-APC (Miltenyi Biotec, cat# 130-111-909, 1:50), MSC Phenotyping Cocktail-PE (CD34, CD14, CD19, CD45, Miltenyi Biotec cat# 130-125-285, dilution according to the manufacturer’s instructions).

Techniques: Flow Cytometry, Marker, Expressing, Staining

Journal: Cells

Article Title: Restoration of BDNF, DARPP32, and D2R Expression Following Intravenous Infusion of Human Immature Dental Pulp Stem Cells in Huntington’s Disease 3-NP Rat Model

doi: 10.3390/cells11101664

Figure Lengend Snippet: Engraftment of hIDPSCs four days after IV administration in the striatum, cortex and subventricular zone (SVZ). ( A ) Images show hIDPSC (within striatum) stained with Vybrant (green arrows) and nuclei stained with propidium iodide (red arrows). Analysis performed at day 9 (D9). ( B , C ) Immunohistochemistry demonstrates positive immunostaining for the anti-hNu antibody (black arrows) observed within the striatum ( b1 – b3 ), SVZ (blue arrows) ( b4 ) and cortex ( c1 – c3 ). B2 and B4 show cells immunolabeled with the anti-hNu antibody in the striatal perivascular area and subventricular zone, respectively. Negative control shows the absence of unspecific labeling of secondary antibody control for anti-human nuclei ( c4 ). Analysis performed at day 9 (D9). Immunofluorescence assay (in confocal microscopy) showing the expression of CD73 ( D ) and CD105 in the rat brain ( E ). Yellow arrows demonstrate cells expressing CD73-positive (d1) and CD105-positive (e1) and, white arrows, cells C73 (d2) and CD105 in smallest magnification (e2). These cells exhibits an usual MSC phenotype (N). Analysis performed at day 35 (D35). Total magnification: 5× ( a1–a4 , b1 , b4 , c1 , e1 ), 10× ( c4 , d2 ), 20× ( c2 ) and 40× ( b2 , b3 , c3 , d1 , e2 ). Scale bars: 5 µm ( a1 – a4 , b2 , b4 , c2–c4 , d1 , d2 , e1 , e2 ), 10 μm ( b3 ).

Article Snippet: After this step, the samples were incubated overnight at 4 °C with primary antibodies anti-human CD73 (ab133582) and -CD105 (ab2529), both from Abcan (Cambridge, UK), at a dilution of 1:100 in PBS.

Techniques: Staining, Immunohistochemistry, Immunostaining, Immunolabeling, Negative Control, Labeling, Control, Immunofluorescence, Confocal Microscopy, Expressing