Journal: Cancer research

Article Title: Focused Ultrasound Delivers Targeted Immune Cells to Metastatic Brain Tumors

doi: 10.1158/0008-5472.CAN-12-2609

Figure Lengend Snippet: Histological quantification of HER2-specific NK-92 cells accumulating at the tumor site. Effector cells were co-localized with CD45 IHC (upper panel) and Prussian blue histochemistry (lower panel) in the three experimental groups, A. HER2-specific NK-92 cells reaching the tumor were quantitatively assessed (mean ± SEM), B. When NK-92 cells were injected prior to BBBD, the number reaching the tumor was significantly higher than if they were injected following or without BBBD (group 3 vs groups 1 and 2: 0.95 ± 0.23 vs 0.09 ± 0.11, 0.21 ± 0.15, p < 0.01). There was no statistical difference between groups 1 and 2. These results are in agreement with the iron-sensitive MR imaging in Figure 4.

Article Snippet: Expression of CD45 was evaluated on a cell smear of targeted cells using mouse anti-human CD45 (RnD systems, Minneapolis, MN USA) and the procedures described above.

Techniques: Injection, Imaging

Journal: Cancer research

Article Title: Focused Ultrasound Delivers Targeted Immune Cells to Metastatic Brain Tumors

doi: 10.1158/0008-5472.CAN-12-2609

Figure Lengend Snippet: FUS causes the translocation of HER2-specific NK-92 cells from the vasculature into the brain and tumor when they are present in the circulation at the time of BBBD. A, CD45 IHC depicting a vessel from which a large number of cells have extravasated and appear to track to the tumor (indicated by the star). B, the corresponding Prussian blue stained section is shown, colocalizing the HER2-specific NK-92 cells. C, a normal capillary adjacent the tumor but within the sonicated region, shows HER2-specific NK-92 cells forced to the adluminal surface of the vessel. FUS results in HER2-specific NK-92 cells circumventing both the BBB and BTB. D, the corresponding Prussian blue section. These cell distributions were seen exclusively in group 3 animals.

Article Snippet: Expression of CD45 was evaluated on a cell smear of targeted cells using mouse anti-human CD45 (RnD systems, Minneapolis, MN USA) and the procedures described above.

Techniques: Translocation Assay, Staining, Sonication

Journal: Cancer immunology research

Article Title: Combined anti-VEGF and anti–CTLA-4 therapy elicits humoral immunity to galectin-1 which is associated with favorable clinical outcomes

doi: 10.1158/2326-6066.CIR-16-0385

Figure Lengend Snippet: Anti-Gal-1 antibodies isolated from an Ipi-Bev-treated patient abrogate Gal-1 binding to CD45. Anti-Gal-1 antibodies were affinity purified from plasma. Biotinylated HAS-Gal-1 (250 ng/ml) was incubated with a commercial anti-Gal-1 polyclonal antibody (Gal-1 Ab) or control antibody (10 μg/ml), enriched endogenous Gal-1 antibodies (Gal-1 Ig) or normal human IgG (1.98 μg/ml) prior to incubation with coated CD45. The binding of HAS-Gal-1 to CD45 was detected with streptavidin-HRP. Sucrose and lactose were added to the reaction at 5 mM. Data are presented as Mean ± SD of 3 experiments.

Article Snippet: Recombinant human CD45 (R&D Systems) was coated onto 96-well plates (25 ng/well) at 4°C overnight.

Techniques: Isolation, Binding Assay, Affinity Purification, Clinical Proteomics, Incubation, Control

Journal: Journal of Nanobiotechnology

Article Title: A near infrared light-triggerable modular formulation for the delivery of small biomolecules

doi: 10.1186/s12951-019-0530-y

Figure Lengend Snippet: Intracellular trafficking of RA1-CB[6] @AuNRs as evaluated by immunocytochemistry. a Schematic representation of the experiment. b Confocal images of leukemic cells incubated with RA1-CB[6] @AuNR-PEG-TRITC. Cells were incubated for 4 h with AuNR (50 µg/mL), washed to remove the non-internalised AuNRs, added new cell culture media, irradiated for 2 min with a 780 nm laser (power: 2 W/cm 2 ) and finally fixed with PFA (4%). The cell membrane was immunostained for CD45, while endosomes were immunostained for EEA1. White scale bar corresponds to 30 µm. c Intracellular localization of RA1-CB[6] @AuNRs before NIR light irradiation was confirmed by a Z-stack scan. Scale bar corresponds to 10 µm. d The co-localization between RA1-CB[6] @AuNR-PEG-TRITC and early endosomes before and after irradiation was quantified by image analyses. Results are average ± SEM ( n = 3). Statistical analysis was performed using a t-test. ***Denotes statistical significance (P < 0.001)

Article Snippet: Cell membrane was stained with monoclonal mouse anti-human CD45 antibody, diluted 1:50 (R&D Systems, MAB1430) and using Alexa Fluor-488 goat anti-mouse igG as secondary antibody, diluted 1:1000 (Life technologies, A11001).

Techniques: Immunocytochemistry, Incubation, Cell Culture, Irradiation, Membrane

Journal: Journal of nanobiotechnology

Article Title: Cerium oxide nanoparticles-carrying human umbilical cord mesenchymal stem cells counteract oxidative damage and facilitate tendon regeneration.

doi: 10.1186/s12951-023-02125-5

Figure Lengend Snippet: Fig. 1 Characterization of hUCMSCs. A Primary hUCMSCs were spindle-shaped under light microscopy. B Flow cytometry analysis showed that 98.5% of cells expressed CD29, 97.4% expressed CD44, 98.6% of cells expressed CD90, and 98.1% of cells expressed CD105. Meanwhile, only 0.48% and 0.28% expressed CD45 and CD34. C Lipid droplets were stained by oil red O after adipogenic differentiation induction of hUCMSCs. Mineralized nodules were stained by Alizarin Red after osteogenic differentiation induction of hUCMSCs. Acid mucopolysaccharide was stained by Alcian Blue after chondrogenic differentiation induction of hUCMSCs

Article Snippet: Briefly, 107 hUCMSCs were subjected to analysis for surface markers CD34 (E-AB-F1143D, Elabscience), CD44 (E-AB-F1100D, Elabscience), CD45 (E-AB-F1137D, Elabscience), CD29 (E-AB-F1049D, Elabscience), CD90 (E-AB-F1167D, Elabscience), and CD105 (E-AB-F1310D, Elabscience) by flow cytometry following the manufacturer’s protocol.

Techniques: Light Microscopy, Flow Cytometry, Staining

Journal: Biomolecules

Article Title: The Role of Doxycycline and IL-17 in Regenerative Potential of Periodontal Ligament Stem Cells: Implications in Periodontitis

doi: 10.3390/biom13101437

Figure Lengend Snippet: Morphology, immunophenotype and differentiation potential of PDLSCs: ( A ) Adherent PDLSCs with fibroblast-like shape grown in GM under standard conditions for 3 days (scale bars: 50 µM); Florescent images of TRITC-conjugated phalloidin labeled F-actin (red) merged with DAPI (4′,6-diamidino-2-phenylindole) nuclear staining (blue) (scale bars: 10 µM). ( B ) Immunophenotypic characteristics of PDLSCs estimated by flow cytometry. Representative histograms present percentages of cells positive (empty peaks) for mesenchymal markers (CD29, CD44, CD73 and CD105) and hematopoietic markers (CD11b, CD235, CD34 and CD45) in comparison to isotype control (shaded peaks). ( C ) Tri-lineage differentiation potential of PDLSCs. Cells were cultured in growth (GM) and differentiation medium (DM). Osteogenic differentiation was determined by positive staining for ALP activity and extracellular matrix mineralization with Alizarin red; scale bars: 50 m; positive staining of proteoglycans by Safranin O confirmed chondrogenic differentiation of PDLSCs; scale bars: 50 m; Oil Red O staining of intracytoplasmic lipid droplets demonstrated adipogenic differentiation; scale bars: 20 µm.

Article Snippet: Cells were then labeled for 30 min at +4 °C in the dark with monoclonal antibodies specific for human antigens including CD45, CD235a, CD90, CD44H, CD73 (all from R&D Systems, Minneapolis, MN, USA), CD-105, CD29 (Invitrogen, Waltham, MA, USA), CD34 (Dako Cytomation, Glostrup, Denmark) and CD11b (Biosource, Camarillo, CA, USA), each of which was conjugated with either FITC or PE.

Techniques: Labeling, Staining, Flow Cytometry, Comparison, Control, Cell Culture, Activity Assay