Journal: Scientific Reports

Article Title: A CXCR4 targeting peptide delivered by silica nanoparticles eliminates migrating cancer stem cells in pancreatic ductal adenocarcinoma

doi: 10.1038/s41598-026-48584-2

Figure Lengend Snippet: BMI1 downstream CXCL12–CXCR4 regulates EMT and stemness. ( A ) Protein–protein interactions of CXCL12 and CXCR4 with relevant factors involved in metastasis (red), stemness (purple), sonic hedgehog signaling (blue), AKT signaling (grey) and NFκB pathway (yellow) (STRING). ( B ) BMI1 gene expression analysis and western blot analysis. GAPDH was used as a loading control. Intensity ratios (IR) calculated against control lane using ImageJ. Cropped blot for clarity. ( C ) Gene expression analysis with genes involved in EMT using qRT-PCR. ( D ) Immunofluorescence quantifications and representative micrographs for indicated cell lines with white arrowheads marking mesenchymal structures of actin filaments stained with Phalloidin (pink) and nucleus stained with DAPI (blue). ( E ) Migration assays towards serum containing media. ( F ) Experimental scheme to evaluate CD133 and CXCR4 surface expression using flow cytometry for Panc354 and MetPO1 (sh_ SCR , sh1_ BMI1 and sh2_ BMI1 ) with (or without) CXCL12. ( G ) Flow cytometry analysis of CD133 + cells and CD133 + CXCR4 + cells in MetPO1 (sh_ SCR , sh1_ BMI1 and sh2_ BMI1 ) treated with (or without) CXCL12. ( H ) Representative cytometry plots for MetPO1 cell line. Error bars represent the standard deviation. n = 3 for all experiments unless otherwise depicted in the datasets. * p < 0.05, ns = not significant.

Article Snippet: Recombinant BMI1 IgG1 (REA438) , APC , Miltenyi Biotec , 130-124-301.

Techniques: Protein-Protein interactions, Gene Expression, Western Blot, Control, Quantitative RT-PCR, Immunofluorescence, Staining, Migration, Expressing, Flow Cytometry, Cytometry, Standard Deviation

Journal: Scientific Reports

Article Title: A CXCR4 targeting peptide delivered by silica nanoparticles eliminates migrating cancer stem cells in pancreatic ductal adenocarcinoma

doi: 10.1038/s41598-026-48584-2

Figure Lengend Snippet: JM#21, most potent EPIX4 derivative to target miCSCs. ( A ) Migration assays towards CXCL12 using Panc354 for EPI-X4 and its derivatives at depicted concentrations. Pre-treatment with EPI-X4, WSCO2, JM#21 and the inactive peptide was applied for 30 min. ( B ) Representative micrographs (10x, DAPI staining) of transwell migration assays in Panc354 cells for the indicated conditions and concentrations. ( C ) Migration assays towards CXCL12 for MetPO1 using JM#21 and the inactive peptide at depicted concentrations. ( D ) Quantifications of percent mesenchymal structures after 15 min and 6 h of CXCL12 treatment in MetPO1 cells. JM#21 pre-treatment was applied for 30 min and representative micrographs with white arrowheads marking mesenchymal structures of actin filaments stained with Phalloidin (pink) and nuclear staining using DAPI (blue). ( E ) Gene expression analysis for indicated cell lines with genes involved in EMT and SHH pathway. ( F ) Gene expression analysis for indicated cell lines with genes involved in stemness. ( G ) Sphere formation assays for 1 st and 2 nd generation of sphere formation. ( H ) Western blot analysis of CADHERIN-1, VIMENTIN, CADHERIN-2, NANOG and BMI1 for indicated cell lines. GAPDH was used as a loading control. Intensity ratios (IR) calculated against control lane using ImageJ. Cropped blot for clarity. ( I ) Experimental design for combination therapy analyzing relapse using JM#21, gemcitabine (labelled as G) and paclitaxel (labelled as P). Quantification of cell viability and representative pictures for clonogenic assays after treatment with JM#21 (10 μM ), gemcitabine (Gem) for indicated concentrations as depicted in experimental design in MetPO1 cell line. ( J ) Flow cytometry for CD133 in Panc354 and MetPO1cells for the indicated treatments shown as fold change. Error bars represent the standard deviation. n = 3 for all experiments unless otherwise depicted in the datasets. * p < 0.05, ns = not significant.

Article Snippet: Recombinant BMI1 IgG1 (REA438) , APC , Miltenyi Biotec , 130-124-301.

Techniques: Migration, Staining, Gene Expression, Western Blot, Control, Flow Cytometry, Standard Deviation

Journal: Regenerative Therapy

Article Title: Increasing robustness of in vitro assay for immnosuppressive effect of mesenchymal stromal/stem cells: The role of inflammatory cytokine production by peripheral blood mononuclear cells

doi: 10.1016/j.reth.2024.12.016

Figure Lengend Snippet: Effect of different donor-derived human peripheral blood mononuclear cells (PBMCs) on the in vitro immunomodulatory response of bone marrow (BM)- mesenchymal stem cells (MSCs). (a) Schematic of the protocol for lymphocyte proliferation assays with mitogen as an immunomodulatory potency assay for MSCs. (b) Proliferation of 10 donor-derived PBMCs upon phytohemagglutinin (PHA; 0.2, 0.5, and 1.0 μg/mL) stimulation was assessed through BrdU uptake. BrdU uptake was indicated by luminescence signal (counts per second, CPS). Two-way ANOVA showed that PBMCs proliferation was significantly affected by the stimulating PHA concentration, the presence or absence of MSCs, and their interaction (p < 0.0001). All values indicate the mean ± standard deviation of mean of three biological replicates (n = 3) (∗P < 0.05, ∗∗P < 0.01, ∗∗∗∗P < 0.0001, two-way analysis of variance, followed by Bonferroni's multiple comparison test). (c) Effect of PBMCs derived from 10 donors on the in vitro immunomodulatory response of BM-MSCs. The rate of suppression by BM-MSCs on PBMC proliferation stimulated with three PHA concentrations (0.2, 0.5, and 1.0 μg/mL) is shown as the immunosuppression rate; CPS(PBMC + MSC)/CPS(PBMC).

Article Snippet: Immortalized human adipose tissue-derived MSCs (I-MSC; JCRB1555 HAdpc-26-E6-Bmi-1-TERT) were obtained from JCRB Cell Bank (Ibaraki, Osaka, Japan).

Techniques: Derivative Assay, In Vitro, Potency Assay, Concentration Assay, Standard Deviation, Comparison

Journal: Regenerative Therapy

Article Title: Increasing robustness of in vitro assay for immnosuppressive effect of mesenchymal stromal/stem cells: The role of inflammatory cytokine production by peripheral blood mononuclear cells

doi: 10.1016/j.reth.2024.12.016

Figure Lengend Snippet: Figure summarizing the flow of the reversed effects of MSCs depending on the cytokine production concentration of PHA-stimulated PBMCs in the immunomodulatory potency assay. The illustration of MSC in the figure was done using BioRender: scientific image and illustration software.

Article Snippet: Immortalized human adipose tissue-derived MSCs (I-MSC; JCRB1555 HAdpc-26-E6-Bmi-1-TERT) were obtained from JCRB Cell Bank (Ibaraki, Osaka, Japan).

Techniques: Concentration Assay, Potency Assay, Software