Journal: Stem Cell Research & Therapy

Article Title: Growth differentiation factor 11 alleviates oxidative stress-induced senescence of endothelial progenitor cells via activating autophagy

doi: 10.1186/s13287-024-03975-y

Figure Lengend Snippet: Rat bone marrow-derived CD34 + VEGFR-2 + EPCs. ( A ) Analysis of CD34 + VEGFR-2 + EPCs by flow cytometry. ( B ) CD34 + VEGFR-2 + EPCs (arrows) in the mononuclear cells isolated from bone marrow. Immunostaining. Scale bar = 25 μm. ( C ) CD31 expression in CD34 + VEGFR-2 + EPCs at 2 weeks after induction with VEGF. Immunostaining. Scale bar = 10 μm. ( D , E ) ALK-4 expression in EPCs demonstrated with immunostaining and Western blotting respectively. Scale bar = 25 μm. ( F ) The statistical result of ALK-4 expression. * P < 0.01 versus young group. n = 3

Article Snippet: The cells were then incubated with mouse anti-rat CD34 and rabbit anti-rat VEGFR-2 antibodies (1:100; Santa Cruz Biotech, Dallas, TX, USA) at 4 o C for 50 min, followed by incubation with Alexa Fluor 647-conjugated goat anti-mouse IgG and Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:200; Jackson, West Grove, PA, USA) at 4 o C for 30 min. CD34 + VEGFR-2 + cells in the mononuclear cells were sorted using a Beckman MoFloTM XDP FACS (fluorescence-activated cell sorter; Beckman Coulter, Fullerton, CA, USA).

Techniques: Derivative Assay, Flow Cytometry, Isolation, Immunostaining, Expressing, Western Blot

Journal: PLoS ONE

Article Title: Antitumor and Angiostatic Activities of the Antimicrobial Peptide Dermaseptin B2

doi: 10.1371/journal.pone.0044351

Figure Lengend Snippet: Treatment of xenografted nude mice was started thirteen days after s.c. PC3 cell injection. Experimental group (n = 8) was treated six times per week p.t. with 100 µL Drs B2 (2.5 mg/kg body weight). Control group (n = 7) was treated six times per week p.t. with 100 µL of PBS. A ) The effect of Drs B2 on tumor size versus time of treatment. After 47 days of treatment the mice were sacrificed, body weight was observed; tumors were isolated, B ) weighted and stored at −80°C. C ) PC3 tumor proliferation was evaluated by Ki67 staining of frozen tissue sections. Proliferation was quantified by image J software analysis of Ki67 positive stained cells on the whole tumor section. The data are mean areas ± SD obtained from 6 control mice and 4 mice treated with Drs B2. Scale bar, 50 µm. * p <0.05 versus control (PBS). D ) Tumor vessel formation was observed with anti CD34 antibodies. Angiogenesis was quantified by image analysis of CD34 positive stained endothelial cells. The data are mean areas ± SD obtained from 6 control mice and 4 mice treated with Drs B2. Scale bar, 50 µm * p <0.05 versus control (PBS).

Article Snippet: Mouse monoclonal anti-CD34 was from Hycult Biotech (The Netherlands), secondary antibody, rabbit anti-rat IgG was obtained from Southern Biotech (Clinisciences; France).

Techniques: Injection, Isolation, Staining, Software

Journal: Journal of vascular research

Article Title: Microvascular endothelial cell responses in vitro and in vivo: modulation by zoledronic acid and paclitaxel?

doi: 10.1159/000313876

Figure Lengend Snippet: Fig. 7. The effect of treatment with zole- dronic acid or paclitaxel on spleen vascu- lature. CD1 nude mice were treated with saline, 100 g/kg i.p. zoledronic acid 20 mg/kg i.v. paclitaxel, alone or in combina- tion. Animals were sacrificed and the spleen isolated and processed for histolog- ical analysis of the vasculature following staining with an antibody to CD34. a Con- trol. b Paclitaxel only. c Zoledronic acid only. d Paclitaxel and zoledronic acid.

Article Snippet: Sections were stained using a rat antibody specific for mouse CD34 (MCA1825CA 1: 50, ABDSerotec, UK) and an IgG2A isotype control, EC and vessel morphology were assessed by an experienced histopathologist (S.S.C.) who was blinded to the treatment groups.

Techniques: Saline, Isolation, Staining

Journal: Neural Regeneration Research

Article Title: A vascular endothelial growth factor–loaded chitosanhyaluronic acid hydrogel scaffold enhances the therapeutic effect of adipose-derived stem cells in the context of stroke

doi: 10.4103/nrr.nrr-d-24-00129

Figure Lengend Snippet: Figure 1 ｜ADSC culture and identity verification, and the effect of VEGF on ADSCs. (A) Representative images of ADSCs on days 3 (i) and 5 (ii) of primary culture, as well as third-passage ADSCs (iii). On day 3, the cells were spindle-shaped, isolated, scattered, and adherent. By day 5, the cells had proliferated rapidly, and the cell colonies had expanded. The morphology of the third-passage cells was uniform, displaying a characteristic fish-like, fibrous, and swirling arrangement. Scale bar: 100 μm. (B) Flow cytometric analysis of CD90, CD29, CD34, and CD45 expression on the cell surface of third-passage ADSCs. (C) CCK-8 analysis of the viability of ADSCs treated with different concentrations of VEGF. Data are expressed as mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, vs. Cont group (one-way analysis of variance with Bonferroni’s post hoc test). (D) Enzyme-linked immunosorbent assay analysis of VEGF, b-FGF, TGF-β, BDNF, and IGF-1 concentration by ADSC cells treated with VEGF (50 ng/mL) or left untreated. Data are expressed as mean ± SEM (n = 12). **P < 0.01, ***P < 0.001, vs. VEGF- group (one-way analysis of variance with Bonferroni’s post hoc test [C] or unpaired t-test [D]). ADSCs: Adipose-derived mesenchymal stem cells; BDNF: brain-derived neurotrophic factor; b-FGF: basic fibroblast growth factor; Cont: control; IGF- 1: insulin-like growth factor-1; TGF-β: transforming growth factor-β; VEGF: vascular endothelial growth factor.

Article Snippet: The following fluorochrome-labeled antibodies were utilized: APC-cy7-CD45 (BD Biosciences, San Jose, CA, USA, Cat# 561586, RRID: AB_10896305), PE-CD34 (RayBiotech, Norcross, GA, USA, Cat# 188-10041-PE, RRID: AB_2315728), FITC-CD90 (BD Biosciences, Cat# 561973, RRID: AB_395588), and BV421-CD29 (BD Biosciences, Cat# 564131, RRID: AB_2738613).

Techniques: Isolation, Expressing, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Derivative Assay, Control

Journal: bioRxiv

Article Title: DiPRO1 dependent transcriptional and epigenetic regulation distinctly controls the fate of muscle and mesenchymal cancer cells

doi: 10.1101/2023.01.08.523169

Figure Lengend Snippet: Antitumor activity of DiPRO1 inhibitors in the Ewing sarcoma (EW) subcutaneous tumor xenograft model. Nude mice received s.c. inoculations of A673 Ewing sarcoma cells and were treated with siDiPRO1/jetPEI® and shDiPRO1/jetPEI® nanocomposites or siCtl/jetPEI® scramble (Ctl) at a dose of 0.5 or 1 mg/kg/d. The initial tumor volume on the day of treatment initiation was V0=168±76 mm3 in the first (EW1; n=7 mice per group) and V0=71±24 mm3 in the second (EW2) and third (EW3) independent experiments (n=5 mice per group). A, Uptake by tumor cells of siDiPRO1/jetPEI®/Cy5 nanocomposites. Internalization was followed in live mice (video; lower panel) treated with a single dose (0.5 mg/kg) of Cy5-coupled anti-DiPRO1 siRNA complexes and in extracted tumors (upper panel) 24 h and 72 h after treatment. Control mice were treated with an equimolar dose of non-targeted siCtl/jetPEI® free of Cy5. B, DiPRO1 depletion efficiency in tumors was verified by RT-qPCR. DiPRO1 mRNA expression levels in three independent experiments were normalized to GAPDH, n=9, mean ± SD. DiPRO1 expression in control tumors was referenced to 100%. Statistical analysis was performed using the t -test. C, Tumor progression was compared between the indicated groups (V n /V 0 ). Results from three independent experiments are presented. Two-way ANOVA test was used for statistical analysis at the indicated time points. D, A nalysis of main endpoints of antitumor activity of the si/hDiPRO1 and control nanomedicines against human Ewing sarcoma xenografts. E, Effect of DiPRO1 inhibition on survival of tumor-bearing mice. Kaplan-Meier curves of overall survival for 43 days (n = 12) from the day of xenograft implantation are shown. Log-Rank test (Mantel-Cox) for each pairwise comparison (treatment vs. control) was applied for statistical analysis. Results from two independent experiments (EW1 and EW2) are combined. F, Assessment of body weight loss (BWL) in mice bearing tumor xenografts. The results of two independent experiments (EW1 and EW2) are combined. G, Immunohistochemical staining of excised tumor tissues at day 12. Representative images of tumor sections stained with hematoxylin/eosin/safranin (HES) (arrows indicate striated muscle), and cells positive for internal and peripheral caspase-3 (Cas-3 periph, indicated by arrows), Ki-67, CD34 and TUNEL are shown. H, Quantitative analysis of positively immunostained cells (%) relative to the total number of tumor cells. Six to eleven fields were selected for counting. Necrotic fields were excluded. C, F: All box-plots and growth curves are displayed as means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, P values are shown between indicated groups and appropriate controls.

Article Snippet: For immunohistochemistry, sections were incubated after heat-induced antigen retrieval with rat anti-mouse CD34 antibody (1/20) (Hycult technologies), rabbit monoclonal anti-Ki67 (1/200) (Neomarkers; LabVision), anti-cleaved caspase 3 (1/100) (Cell Signaling), visualized using the polink anti rat kit (GBI Labs) and the peroxidase/diaminobenzidine Rabbit PowerVision kit (ImmunoVision Technologies), respectively.

Techniques: Activity Assay, Control, Quantitative RT-PCR, Expressing, Inhibition, Comparison, Immunohistochemical staining, Staining, TUNEL Assay

Journal: Molecular medicine reports

Article Title: Interleukin-17 potently increases non-small cell lung cancer growth.

doi: 10.3892/mmr.2015.4694

Figure Lengend Snippet: Figure 2. CD34 and CCR2 expression in F4/80-positive macrophages in implanted tumor masses analyzed by flow cytometry. At day 14, 16 and 18 after implantation, cells were obtained from the implanted tumor masses by dissection. (A) Cells were homogenized, stained with CD34 antibodies and examined by flow cytometry. (B) Quantification of results. A significantly increased proportion of CD34 in the IL-17 group compared with control. (C) Cells were stained with CCR2 antibodies and analyzed by flow cytometry. (D) CCR2-positive macrophages were increased in the IL-17 group compared with control. (*P<0.05, compared with control). IL, interleukin.

Article Snippet: Rat anti-mouse CD34 monoclonal antibody (mAb) (MEC 14.7, cat no. NB600-1071), phycoerythrin (PE)-conjugated rat antimouse F4/80 mAb (cat no. NBP2-22134), rat anti-mouse vascular endothelial growth factor (VEGF) mAb (RM0009-2G02, cat no. NB110-61025), rabbit anti-mouse matrix metalloproteinase (MMP)2 mAb (cat no. NB200-193), rabbit anti-mouse MMP9 mAb (cat no. NBP1-39597), rabbit anti-mouse tumor necrosis factor (TNF)-α mAb (cat no. NB100-57620) and mouse antimouse glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mAb (cat no. NB300-328H) were purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing, Flow Cytometry, Dissection, Staining, Control

Journal: Circulation journal : official journal of the Japanese Circulation Society

Article Title: Hypoxic preconditioning enhances angiogenic potential of bone marrow cells with aging-related functional impairment.

doi: 10.1253/circj.cj-11-0605

Figure Lengend Snippet: Figure 1. Characterization of bone marrow cells (BMCs). Freshly isolat- ed BMCs from young and aged mice were used for flow cytometric analy- sis (A,B), western blot analysis (C), and telomeric repeat amplification protocol assay (D). The numbers of CD34 (A) and c-kit (B) positive stem cells in BMCs were lower in aged mice than in young mice. The expres- sion of p53 was higher (C) and telom- erase activity was lower (D) in BMCs from aged mice. *P<0.05, **P<0.01 and ***P<0.001 vs. cells from young mice (n=3 or 4).

Article Snippet: Briefly, cells were stained with a fluorescein isothiocyanate (FITC)-conjugated rat anti-mouse CD34 antibody (RAM34, eBioscience Inc, San Diego, CA, USA) or a phycoerythrin (PE)-conjugated rat anti-mouse c-kit antibody (2B8, eBioscience Inc).

Techniques: Western Blot, Amplification, Activity Assay