Journal: World Journal of Gastrointestinal Oncology

Article Title: Mesenchymal stem cell-derived lncRNAs NKILA contributes to stemness and chemoresistance by fatty acid oxidation in gastric cancer via miR-485-5p/STAT3

doi: 10.4251/wjgo.v17.i8.105006

Figure Lengend Snippet: Mesenchymal stromal cells enhances stemness and chemoresistance of gastric cancer cells. A-E: The MKN45 and AGS cells were co-cultured with mesenchymal stromal cells (MSCs); The stemness was analyzed using sphere formation assay (A); The mRNA expression of SOX2, Oct4, LIN28, and CD133 was measured by qPCR (B); The levels of SOX2, Oct4, LIN28, and CD133 was detected by Western blot analysis (C); The CD44+ and CD133+ cells were identified by flow cytometry analysis (D and E); F and G: The MKN45 and AGS cells were co-cultured with MSCs and treated with oxaliplatin or 5-Fu; The cell apoptosis was determined by flow cytometry analysis (F); The cell proliferation was examined by colony formation assay (G). a P < 0.01; MSC: Mesenchymal stromal cell.

Article Snippet: Primary human bone marrow-derived MSCs (HUXMA-01001, Procell, China) were cultured in α-MEM medium (Hyclone, United States) supplemented with 10% FBS and 1% penicillin/streptomycin.

Techniques: Cell Culture, Tube Formation Assay, Expressing, Western Blot, Flow Cytometry, Colony Assay

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: Primary human bone marrow-derived MSCs (BM-MSC) were purchased from Lonza (Walkersville, MD, USA) (PT-2501).

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: Relationship between the production of cytokines by PHA-stimulated PBMCs and the suppression rate of PBMC proliferation by MSCs in the immunomodulatory potency assay. Spearman correlation analysis between the concentration of each cytokine in the culture supernatant when PBMCs were stimulated with 1.0 μg/mL PHA and the rate of proliferation inhibition by BM-MSCs in the immunomodulatory potency assay is shown with linear regression lines and their 95 % confidential intervals. r, correlation coefficient.

Article Snippet: Primary human bone marrow-derived MSCs (BM-MSC) were purchased from Lonza (Walkersville, MD, USA) (PT-2501).

Techniques: Potency Assay, Concentration Assay, Inhibition

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: Inter-test variability in the rate of proliferation inhibition of PBMCs by repeated one-way MLR assay.

Article Snippet: Primary human bone marrow-derived MSCs (BM-MSC) were purchased from Lonza (Walkersville, MD, USA) (PT-2501).

Techniques: Inhibition, Mlr Assay

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: Primary human bone marrow-derived MSCs (BM-MSC) were purchased from Lonza (Walkersville, MD, USA) (PT-2501).

Techniques: Concentration Assay, Potency Assay, Software

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Functionality of lyophilized osteoinductive EVs: a mechanistic study

doi: 10.3389/fbioe.2024.1452428

Figure Lengend Snippet: Endocytosis of EVs by HMSCs (A) Representative confocal micrograph of fluorescently labeled fresh and lyophilized BMP2 EVs (green) endocytosed by naïve HMSCs at 37°C. Scale bar = 20 μm. (B) Dose-dependent endocytosis of fresh and lyophilized BMP2 EVs by HMSCs. Data points represent mean percentage fluorescence ± SD (n = 6). (C) Graph showing the inhibition of EV endocytosis after pre-treatment of the EVs with heparin to block interaction with the cell surface HSPGs. The reduction of EV endocytosis at 4°C was also measured compared to 37°C. Data represent mean percentage fluorescence with respect to control ± SD (n = 6). *: statistical significance with respect to No blocking control, #: statistical significance with respect to BMP2 EV group as measured by Student’s t-test (P< 0.05).

Article Snippet: Primary human bone marrow derived MSCs (HMSCs) were purchased from Lonza and cultured in αMEM basal media (Gibco) supplemented with 20% fetal bovine serum (FBS, Gibco), 1% (v/v) L-Glutamine (Gibco) and 1% (v/v) antibiotic-antimycotic solution (Gibco).

Techniques: Labeling, Fluorescence, Inhibition, Blocking Assay, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Functionality of lyophilized osteoinductive EVs: a mechanistic study

doi: 10.3389/fbioe.2024.1452428

Figure Lengend Snippet: Osteoinductive property of the lyophilized BMP2 EVs (A) Osteoinductive gene expression levels in fresh and lyophilized BMP2 EV HMSCs at day 3 and 5 with respect to control (no EV) group. Data are represented as mean fold changes ± SD (n = 4). *: statistical significance with respect to BMP2 EV group as measured by student’s t-test (P< 0.05). (B) Graph representing relative luciferase activity in HMSCs transfected with a BMP2 reporter luciferase plasmid (n = 4). Recombinant BMP2 protein (BMP2 GF) was used as positive control. Note the significant increase in luciferase activity in fresh and lyophilized BMP2 EV treated groups compared to control. *: statistical significance with respect to control, #: statistical significance with respect to BMP2 GF group as measured by Tukey’s test post ANOVA (P< 0.05). (C) Graph representing relative alkaline phosphatase (ALP) activity in HMSCs treated with osteogenic differentiation medium (OS) in the presence/absence of fresh and lyophilized BMP2 EVs at 3 and 7 days (n = 4). *: statistical significance with respect to control, #: statistical significance with respect to OS + BMP2 EV group as measured by Student’s t-test (P< 0.05). (D) Quantitative in-cell western for the presence of phosphorylated SMAD 1/5/8 and the corresponding tubulin expression in a 96 well plate assay (n = 6). The graph represents quantitation of the fluorescence from the wells using the LICOR Odyssey imager normalized to tubulin. *: statistical significance with respect to control group as measured by Tukey’s test post ANOVA (P< 0.05).

Article Snippet: Primary human bone marrow derived MSCs (HMSCs) were purchased from Lonza and cultured in αMEM basal media (Gibco) supplemented with 20% fetal bovine serum (FBS, Gibco), 1% (v/v) L-Glutamine (Gibco) and 1% (v/v) antibiotic-antimycotic solution (Gibco).

Techniques: Gene Expression, Control, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Recombinant, Positive Control, In-Cell ELISA, Expressing, Quantitation Assay, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Functionality of lyophilized osteoinductive EVs: a mechanistic study

doi: 10.3389/fbioe.2024.1452428

Figure Lengend Snippet: Binding of lyophilized EVs to ECM proteins (A) Representative confocal image showing the binding of fluorescently labeled lyophilized EVs (green) to the decellularized ECM of HMSCs. Counterstain was performed for type I collagen (red, scale bar = 5 μm). (B) Dose-dependent binding of fluorescently labeled lyophilized EVs to type I collagen coated assay plates (data points represent mean ± SD, n = 6). (C) Representative confocal image showing the binding of fluorescently labeled lyophilized EVs (green) to the decellularized ECM of HMSCs. Counterstain was performed for fibronectin (red, scale bar = 20 μm). (D) Dose-dependent binding of fluorescently labeled lyophilized EVs to fibronectin coated assay plates (data points represent mean ± SD, n = 6).

Article Snippet: Primary human bone marrow derived MSCs (HMSCs) were purchased from Lonza and cultured in αMEM basal media (Gibco) supplemented with 20% fetal bovine serum (FBS, Gibco), 1% (v/v) L-Glutamine (Gibco) and 1% (v/v) antibiotic-antimycotic solution (Gibco).

Techniques: Binding Assay, Labeling