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: The American Journal of Pathology

Article Title: Identification and Characterization of Mesenchymal-Epithelial Progenitor-Like Cells in Normal and Injured Rat Liver

doi: 10.1016/j.ajpath.2014.08.029

Figure Lengend Snippet: Primary Antibodies Used in This Study

Article Snippet: Mouse anti-rat NT5E , IgG1 , BD Biosciences , 551123 , 1:50.

Techniques: Transduction

Journal: The American Journal of Pathology

Article Title: Identification and Characterization of Mesenchymal-Epithelial Progenitor-Like Cells in Normal and Injured Rat Liver

doi: 10.1016/j.ajpath.2014.08.029

Figure Lengend Snippet: Immunofluorescence microscopy analysis for expression of mesenchymal and epithelial cell marker proteins in 13-11-3-2 cell line. Mesenchymal cell markers were as follows: vascular cell adhesion molecule 1 (VCAM; A); Desmin (B); ACTA2 (α smooth muscle actin; C); glial fibrillary acidic protein (GFAP; F); Vimentin (G); Thy1 (CD90; H); CD73 (Ecto-5′-Nucleotidase; K); and CD29 (Integrin β1; L); Epithelial cell markers were as follows: KRT18 (Cytokeratin 18; D); OV-6 (E); CD44 (I); CDH1 (epithelial cadherin; J); CLDN7 (Claudin-7; M); phase-contrast microphotograph of cultured 13-11-3-5 cells (N); and phase-contrast microphotograph of cultured 13-11-3-3 cells (O). Original magnifications: ×40 (M); ×10 (N and O).

Article Snippet: Mouse anti-rat NT5E , IgG1 , BD Biosciences , 551123 , 1:50.

Techniques: Immunofluorescence, Microscopy, Expressing, Marker, Cell Culture

Journal: BMC Biotechnology

Article Title: Layer-shaped alginate hydrogels enhance the biological performance of human adipose-derived stem cells

doi: 10.1186/1472-6750-12-35

Figure Lengend Snippet: Immunophenotypic analysis of the third passage hADSCs by flow cytometry. The fluorescence level detected for the stromal stem cells markers was 95.9% for CD44, 92.04% for CD73, 92.86% for CD90 and 94.6% for CD105, respectively, whereas for the hematopoietic stem cells marker CD34 only 0.3% of the gated cells were fluorescently labeled. The positive zones in the histograms were marked with B in the fluorescence channel 1 (FL1), and with C in the fluorescence channel 2 (FL2), respectively, based on the isotype control antibody.

Article Snippet: We examined the expression of the following cell surface antigens: CD34 (mouse anti-human-PE conjugated monoclonal antibody, code A07776, Beckman Coulter) as a hematopoietic marker and CD44 (mouse anti-human monoclonal antibody, 1:50, code Sc-9960, Santa-Cruz Biotechnology), CD73 (rabbit anti-human polyclonal antibody, 1:50, code Sc-25603, Santa-Cruz Biotechnology), CD90 (mouse anti-human monoclonal antibody, 1:50, code Sc-59396, Santa-Cruz Biotechnology) and CD105 (mouse anti-human-PE conjugated monoclonal antibody, code A07414, Beckman Coulter) as typical protein markers in MSCs.

Techniques: Flow Cytometry, Fluorescence, Marker, Labeling, Control

Journal: Scientific Reports

Article Title: PET imaging of colon cancer CD73 expression using cysteine site-specific 89 Zr-labeled anti-CD73 antibody

doi: 10.1038/s41598-024-68987-3

Figure Lengend Snippet: Cysteine-site specific 89 Zr labeling of anti-CD73 IgG. ( A ) Non-reduced SDS-PAGE of unmodified, TCEP-reduced, and DFO-conjugated anti-CD73 IgG. The Amicon filtered lane refers to DFO-conjugated antibody following Amicon filtration to remove unconjugated reagents. ( B ) Autoradiography of peak fractions of 89 Zr-CD73 IgG by native PAGE (right). ( C ) Radioactivity profile of PD-10 column-eluted fractions. ( D ) In vitro stability of 89 Zr-CD73 IgG assessed by radio-iTLC.

Article Snippet: Rat IgG2a against mouse CD73 (clone TY/23; BioXcell, West Lebanon, NH) and isotype IgG2a (clone 2A3; BioXcell) were site-specifically conjugated with deferoxamine–maleimide on sulfhydryl residues as previously described .

Techniques: Labeling, SDS Page, Filtration, Autoradiography, Clear Native PAGE, Radioactivity, In Vitro

Journal: Scientific Reports

Article Title: PET imaging of colon cancer CD73 expression using cysteine site-specific 89 Zr-labeled anti-CD73 antibody

doi: 10.1038/s41598-024-68987-3

Figure Lengend Snippet: CD73 expression and 89 Zr-CD73 IgG binding in colon cancer cells. ( A ) ( a ) Immunoblotting of CD73 in low-expressing parental CT26 and high-expressing CT26/CD73 cancer cells. ( b ) 89 Zr-CD73 IgG binding in CT26, CT26/CD73 cells, and in CT26/CD73 cells blocked with excess unlabeled anti-CD73 antibody. ( B ) The CD73 protein amount ( a ) and 89 Zr-CD73 IgG uptake level, ( b ) are increased proportionally to the CT26/CD73 cell content in a mixture of CT26/CD73 and CT26 cells. ( C ) Representative Lindmo binding assay. A conventional plot of specific and non-specific binding over total applied radioactivity, as a function of increasing cell concentration, is shown ( a ). A double inverse plot was drawn using the same data as those describing total applied radioactivity over specific binding, as a function of the inverse cell concentration ( b ). The immunoreactive fraction was determined through linear extrapolation to the ordinate. All data are the mean ± standard deviation values obtained from a single experiment (n = 3 per group).

Article Snippet: Rat IgG2a against mouse CD73 (clone TY/23; BioXcell, West Lebanon, NH) and isotype IgG2a (clone 2A3; BioXcell) were site-specifically conjugated with deferoxamine–maleimide on sulfhydryl residues as previously described .

Techniques: Expressing, Binding Assay, Western Blot, Radioactivity, Concentration Assay, Standard Deviation

Journal: Scientific Reports

Article Title: PET imaging of colon cancer CD73 expression using cysteine site-specific 89 Zr-labeled anti-CD73 antibody

doi: 10.1038/s41598-024-68987-3

Figure Lengend Snippet: CD73 expression and 89 Zr-CD73 IgG binding in 4T1.2 breast cancer cells. ( A ) Immunoblotting and quantified band intensities of CD73 protein in 4T1.2 cells compared to overexpressing CT26/CD73 cells. ( B ) 89 Zr-CD73 IgG binding in 4T1.2 cells compared to equal number of CT26/CD73 cells, and effect of blocking with excess unlabeled anti-CD73 antibody. All data are the mean ± standard deviation values obtained from a single experiment (n = 3 per group).

Article Snippet: Rat IgG2a against mouse CD73 (clone TY/23; BioXcell, West Lebanon, NH) and isotype IgG2a (clone 2A3; BioXcell) were site-specifically conjugated with deferoxamine–maleimide on sulfhydryl residues as previously described .

Techniques: Expressing, Binding Assay, Western Blot, Blocking Assay, Standard Deviation

Journal: Scientific Reports

Article Title: PET imaging of colon cancer CD73 expression using cysteine site-specific 89 Zr-labeled anti-CD73 antibody

doi: 10.1038/s41598-024-68987-3

Figure Lengend Snippet: 89 Zr-CD73 IgG PET/CT and biodistribution at 4 days in CT26 and CT26/CD73 tumor mice. ( A ) Coronal and transaxial tomographic PET images at 4 days post-injection showing high 89 Zr-CD73 IgG uptake in CT26/CD73 tumors (arrow). CD73-specific uptake was verified by low 89 Zr-CD73 IgG accumulation in CT26 tumors. Target specificity was further supported by low CT26/CD73 tumor-contrast in a separate control group injected with 89 Zr-isotype IgG. ( B ) Biodistribution data reiterated these findings by confirming greater 89 Zr-CD73 IgG uptake in CT26/CD73 tumors compared to CT26 tumors. ( C ) Furthermore, the CT26/CD73 tumor-to-blood ratio was significantly higher for 89 Zr-CD73 IgG compared to 89 Zr-isotype IgG. Data are presented as the mean ± standard deviation values obtained from a single experiment (n = 5 per group).

Article Snippet: Rat IgG2a against mouse CD73 (clone TY/23; BioXcell, West Lebanon, NH) and isotype IgG2a (clone 2A3; BioXcell) were site-specifically conjugated with deferoxamine–maleimide on sulfhydryl residues as previously described .

Techniques: Positron Emission Tomography-Computed Tomography, Injection, Control, Standard Deviation

Journal: Scientific Reports

Article Title: PET imaging of colon cancer CD73 expression using cysteine site-specific 89 Zr-labeled anti-CD73 antibody

doi: 10.1038/s41598-024-68987-3

Figure Lengend Snippet: 89 Zr-CD73 IgG PET/CT and biodistribution at 8 days in CT26 and CT26/CD73 tumor mice. ( A ) Coronal tomographic and maximum intensity–projection (MIP) 89 Zr-CD73 IgG PET/CT in CT26 (left) or CT26/CD73 tumor-bearing mice (middle), and 89 Zr-isotype IgG PET/CT in CT26/CD73 tumor mice (right). ( B ) Biodistribution data of mice as above at 8 days post-injection. (C) Again, the CT26/CD73 tumor-to-blood ratio was significantly higher for 89 Zr-CD73 IgG compared to 89 Zr-isotype IgG. Data are the mean ± standard error of obtained from two independent experiments (n = 5 per group).

Article Snippet: Rat IgG2a against mouse CD73 (clone TY/23; BioXcell, West Lebanon, NH) and isotype IgG2a (clone 2A3; BioXcell) were site-specifically conjugated with deferoxamine–maleimide on sulfhydryl residues as previously described .

Techniques: Positron Emission Tomography-Computed Tomography, Injection

Journal: Scientific Reports

Article Title: PET imaging of colon cancer CD73 expression using cysteine site-specific 89 Zr-labeled anti-CD73 antibody

doi: 10.1038/s41598-024-68987-3

Figure Lengend Snippet: CD73 expression in CT26 and CT26/CD73 tumor tissues. ( A ) Immunoblotting for CD73 and β-actin. Note that CD73 bands in CT26 tumors were detected only by long exposure times (middle) while they were detected in CT26/CD73 tumors by short exposure times (top). ( B ) Immunohistochemistry showing strong CD73 staining in CT26/CD73 tumors ( a ) and weak staining in CT26 tumors ( b ). Arrows point to tumor cells with positive CD73 staining. The area in the box for the lower magnified images (top; ×40) are shown in the higher magnified images (bottom ×100).

Article Snippet: Rat IgG2a against mouse CD73 (clone TY/23; BioXcell, West Lebanon, NH) and isotype IgG2a (clone 2A3; BioXcell) were site-specifically conjugated with deferoxamine–maleimide on sulfhydryl residues as previously described .

Techniques: Expressing, Western Blot, Immunohistochemistry, Staining

Journal: Scientific Reports

Article Title: PET imaging of colon cancer CD73 expression using cysteine site-specific 89 Zr-labeled anti-CD73 antibody

doi: 10.1038/s41598-024-68987-3

Figure Lengend Snippet: 89 Zr-CD73 IgG PET/CT and biodistribution at 8 days in 4T1.2 tumor mice. ( A ) Coronal tomographic and maximum intensity-projection (MIP) 89 Zr-CD73 (left) and 89 Zr-isotype IgG (middle) PET/CT in mice bearing CT26/CD73 tumors. Transaxial tomographs are shown in the right. ( B ) Biodistribution data of mice as above at 8 days post-injection. ( C ) Again, the CT26/CD73 tumor-to-blood ratio was significantly higher for 89 Zr-CD73 IgG compared to 89 Zr-isotype IgG. Data are presented as the mean ± standard error of obtained from two independent experiments (n = 5 per group).

Article Snippet: Rat IgG2a against mouse CD73 (clone TY/23; BioXcell, West Lebanon, NH) and isotype IgG2a (clone 2A3; BioXcell) were site-specifically conjugated with deferoxamine–maleimide on sulfhydryl residues as previously described .

Techniques: Positron Emission Tomography-Computed Tomography, Injection

Journal: International journal of molecular sciences

Article Title: Combination of Epigallocatechin-3-Gallate and Tramiprosate Prevent Accumulation of Intracellular Aβ and Dysfunctional Autophagy-Lysosomal Pathway at Earliest Stage of Transdifferentiation of Mesenchymal Stromal Cells into PSEN1 E280A Cholinergic-like Neurons.

doi: 10.3390/ijms26083756

Figure Lengend Snippet: Figure 3. Flow cytometry and immunofluorescence analysis of proliferation marker Ki67 and cluster of differentiation 73 (CD73) during the process of transdifferentiation of menstrual mesenchymal stromal cells (MenSCs) into cholinergic-like neurons (ChLNs). Representative histograms showing Ki67 (A–E) and CD73 (F–J) flow cytometry analysis performed in wild-type (WT) PSEN1 (blue his- tograms) and E280A (red histograms) MenSCs after 0 (A,F), 1 (B,G), 3 (C,H), 5 (D,I), and 7 (E,J) days of culture in Ch-N-Rm. (K,L) Bar charts represent the quantitative analysis of the positive population. Representative images showing Ki67 (green) and CD73 (red) double-stained WT PSEN1 (M-Q) and E280A (R–V) MenSCs after 0 (M,R), 1 (N,S), 3 (O,T), 5 (P,U), and 7 (Q,V) days of culture in Ch-N-Rm. Nuclei were stained with Hoechst dye (blue; M–V). (W,X) Bar charts represent the mean fluorescent intensity (MFI) quantification of images obtained by immunofluorescence analysis. The histograms and images represent one out of three independent experiments. Significant values were determined by two-way ANOVA with a Tukey post hoc test; ** p < 0.01, *** p < 0.001 intergroup comparison; n.s. = not significant. Image magnification 20×.

Article Snippet: For the analysis of differentiation, neuronal, Alzheimer’s disease, oxidative stress, autophagy, and cell death-related markers, cells treated under different conditions were washed and incubated with 10% bovine serum albumin (BSA) blocking solution for 30 min. For the analysis of neuronal Alzheimer’s disease, oxidative stress, autophagy, and cell death-related markers, cells were incubated overnight with primary antibodies against Ki-67 (rabbit; abcam; cat. #ab15580, Cambridge, UK), CD73 (mouse; Elabscience; cat. #E-AB-F1242E, Houston, TX, USA), NeuN (rabbit; abcam; cat. #ab177487, Cambridge, UK), ChAT (ChAT, Millipore; cat. #AB144 P, Burlington, MA, USA), protein amyloid β1-42 (mouse, Millipore; cat. #MABN639, Burlington, MA, USA), oxidized DJ-1 (oxDJ-1; spanning residue C106 of human PARK7/DJ1; oxidized to cysteine sulfonic acid (SO3); rabbit, abcam; cat. #ab169520, Cambridge, UK), and LC3-II (rabbit, Novusbio; cat. #NB100-2220, Centennial, Colorado, USA) or 1X CellEvent Caspase3/7 Detection Reagent (Invitrogen, cat. # C10423, Carlsbad, CA, USA).

Techniques: Flow Cytometry, Immunofluorescence, Marker, Staining, Comparison

Journal: Toxicology Reports

Article Title: In utero arsenic exposure increases DNA damage and gene expression changes in umbilical cord mesenchymal stem cells (UC-MSCs) from newborns as well as in UC-MSC differentiated hepatocytes

doi: 10.1016/j.toxrep.2022.09.002

Figure Lengend Snippet: Immunohistochemical detection of MSC markers and DNA damage (8-OHdG and 8-nitroguanine) in paraffin-embedded umbilical cord sections. Paraffin-embedded sections of umbilical cord samples from 100 arsenic-exposed newborns were subjected to immunohistochemical detection of MSC markers and 8-OHdG and 8-nitroguanine. The drawing picture of umbilical cord composition and the staining procedure for MSC markers and 8-OHdG and 8-nitroguanine in umbilical cord cross-sections were modified from a study by Shugar et al. (A) (WJ: Wharton’s jelly). Representative images showing localization of MSC markers in Wharton’s jelly; CD73 (red), CD90 (yellow), and CD105 (orange) in umbilical cord tissue as judged by co-localization of DAPI (blue) as indicated in merge (white) (B-upper). Representative images showing localization of 8-OHdG (green), and 8-nitroguanine (pink) in umbilical cord MSCs as judged by co-localization of DAPI (blue) as indicated in merge (white) (B-lower). Cells were viewed at 200× magnification.

Article Snippet: Secondly, Alexa Fluor®750 goat immunoglobulin G (IgG) anti-rabbit secondary antibody (1:500, Abcam, UK), Alexa Fluor®594 goat IgG anti-mouse secondary antibody (1:500, Molecular Probes, USA), mouse monoclonal anti-human CD73 conjugated to PE (1:50, Abcam), mouse monoclonal anti-human CD105 conjugated to PerCP-CyTM5.5 (1:50, BD PharmigenTM, USA), and mouse monoclonal anti-8-OHdG conjugated to Alexa Fluor®488 (1:100, Santa Cruze Biotechnology, USA) were incubated for 1 h in the dark at 37 °C.

Techniques: Immunohistochemical staining, Staining, Modification