Journal: Cytotherapy

Article Title: Mesenchymal stromal cell delivery of full-length tumor necrosis factor–related apoptosis-inducing ligand is superior to soluble type for cancer therapy

doi: 10.1016/j.jcyt.2015.03.603

Figure Lengend Snippet: Expression of recombinant TRAIL by transduced MSCs. (A) Schematic description of TRAIL expression constructs. (B) Fluorescence-activated cell sorting analysis of lentivirus-transduced MSCs. (C) Long-term fluorescence-activated cell sorting analysis of TRAIL expression in MSCs transduced at passage 3 (p3), expanded and passaged every 7 days until p8. (D) Detection of TRAIL and α-tubulin expression by immunoblotting. Cont. represents MSC-GFP cell lysates or supernatant medium; flT and sT represent cell lysates or concentrated culture supernatant from MSC-flT and MSC-sT cells, respectively; rT represents 1 ng of purified recombinant human TRAIL (amino acids 114–281) produced from bacterial cells (PeproTech). (E) Levels of TRAIL in cell culture supernatants from MSCs transduced at p3 and expanded for one passage, measured by ELISA. Data presented as TRAIL released by 1 million cells per hour (ng/h/1 × 10 6 cells). Data represent averages ± SEM ( n = 5).

Article Snippet: DiI-labeled cancer cells ( n = 8000) were plated into one well of a 96-well plate, to which transduced MSCs, purified recombinant TRAIL 50 ng/mL (amino acids 114–281) (Peprotech), the pan-caspase inhibitor Z-VAD-FMK (1 μg/mL [Sigma]), a neutralizing monoclonal anti-TRAIL Ab (10 ng/mL [Sigma, Cat. No. T3067]) or control medium was added for 24 h. Ratios of MSCs to cancer cells included 1:10, 2:10, 4:10, 6:10, 8:10 and 10:10 (MSC: cancer cell).

Techniques: Expressing, Recombinant, Construct, Fluorescence, FACS, Western Blot, Purification, Produced, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Cytotherapy

Article Title: Mesenchymal stromal cell delivery of full-length tumor necrosis factor–related apoptosis-inducing ligand is superior to soluble type for cancer therapy

doi: 10.1016/j.jcyt.2015.03.603

Figure Lengend Snippet: Immunofluorescent detection of recombinant TRAIL (red) expressed by transduced MSCs. Phalloidin staining was used to show filamentous actin (green); nuclei were labeled with DAPI (blue). Top panel shows intracellular staining; bottom panel shows cell surface staining. Images are representative of at least three experiments for each staining condition. Magnification ×400.

Article Snippet: DiI-labeled cancer cells ( n = 8000) were plated into one well of a 96-well plate, to which transduced MSCs, purified recombinant TRAIL 50 ng/mL (amino acids 114–281) (Peprotech), the pan-caspase inhibitor Z-VAD-FMK (1 μg/mL [Sigma]), a neutralizing monoclonal anti-TRAIL Ab (10 ng/mL [Sigma, Cat. No. T3067]) or control medium was added for 24 h. Ratios of MSCs to cancer cells included 1:10, 2:10, 4:10, 6:10, 8:10 and 10:10 (MSC: cancer cell).

Techniques: Recombinant, Staining, Labeling

Journal: Cytotherapy

Article Title: Mesenchymal stromal cell delivery of full-length tumor necrosis factor–related apoptosis-inducing ligand is superior to soluble type for cancer therapy

doi: 10.1016/j.jcyt.2015.03.603

Figure Lengend Snippet: MSC-flT cells induce cancer cell apoptosis with a higher efficiency than MSC-sT cells. (A–D) Apoptosis of five highly TRAIL-sensitive cancer cell lines, Colo205, NCI-H460, H727, H2795 and H2804 (A); five moderately TRAIL-sensitive cancer cell lines, H2731, H226, H2869, PC9 and M231 (B); four cancer cell lines of low TRAIL sensitivity, HT29, H357, H2452 and RKO (C); and six TRAIL-resistant cancer cell lines, A549, NCI-H2052, H2810, NCI-H23, RCC10 and HA7-RCC (D) was determined after 24 h of culture with medium (control), 50 ng/mL of purified recombinant TRAIL (rTRAIL), MSC-GFP, MSC-flT or MSC-sT, with a ratio of 4 p4 MSC to 10 cancer cells. Data represent averages of three experiments with triplicate repeats for each cell line.

Article Snippet: DiI-labeled cancer cells ( n = 8000) were plated into one well of a 96-well plate, to which transduced MSCs, purified recombinant TRAIL 50 ng/mL (amino acids 114–281) (Peprotech), the pan-caspase inhibitor Z-VAD-FMK (1 μg/mL [Sigma]), a neutralizing monoclonal anti-TRAIL Ab (10 ng/mL [Sigma, Cat. No. T3067]) or control medium was added for 24 h. Ratios of MSCs to cancer cells included 1:10, 2:10, 4:10, 6:10, 8:10 and 10:10 (MSC: cancer cell).

Techniques: Control, Purification, Recombinant

Journal: Cytotherapy

Article Title: Mesenchymal stromal cell delivery of full-length tumor necrosis factor–related apoptosis-inducing ligand is superior to soluble type for cancer therapy

doi: 10.1016/j.jcyt.2015.03.603

Figure Lengend Snippet: MSC-flT–derived cell surface and secreted TRAILs contribute to induction of apoptosis in cancer cells. (A, B) Apoptosis was measured by means of flow cytometry after exposure of M231 cells (A) or A549 cells (B) for 24 h to increasing doses of recombinant TRAIL (rTRAIL), supernatant TRAIL from MSC-sT (sTRAIL) or supernatant TRAIL from MSC-flT (flTRAIL). (C) Apoptosis was measured after 24-h co-culture of A549 cells with live or fixed MSC-GFP and MSC-flT cells, respectively. Data represent averages ± SEM ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01 compared with rTRAIL and MSC-GFP treatment, respectively, by Student's t -test.

Article Snippet: DiI-labeled cancer cells ( n = 8000) were plated into one well of a 96-well plate, to which transduced MSCs, purified recombinant TRAIL 50 ng/mL (amino acids 114–281) (Peprotech), the pan-caspase inhibitor Z-VAD-FMK (1 μg/mL [Sigma]), a neutralizing monoclonal anti-TRAIL Ab (10 ng/mL [Sigma, Cat. No. T3067]) or control medium was added for 24 h. Ratios of MSCs to cancer cells included 1:10, 2:10, 4:10, 6:10, 8:10 and 10:10 (MSC: cancer cell).

Techniques: Derivative Assay, Flow Cytometry, Recombinant, Co-Culture Assay

Journal: Stem Cells and Cloning : Advances and Applications

Article Title: Characterization of neural stemness status through the neurogenesis process for bone marrow mesenchymal stem cells

doi: 10.2147/SCCAA.S94545

Figure Lengend Snippet: Morphology characteristics of mouse MSCs cultured in MEM +20% FBS after 24 hours as revealed under inverted microscope. Notes: ( A ) 10 x ; ( B ) 20x. Abbreviations: MSCs, mesenchymal stem cells; MEM, minimum essential medium; FBS, fetal bovine serum.

Article Snippet: There were four different formulas: 1) MEM media with 5% FBS alone, 2) MEM supplemented with 50 ng/mL epidermal growth factors (EGFs; United States Biological, MA, USA; E3374-11), 3) MEM supplemented with 50 ng/mL basic fibroblast growth factors (b-FGFs) (United States Biological; F4210-20M1), and 4) MEM with 50 ng/mL for each b-FGF and EGF.

Techniques: Cell Culture, Inverted Microscopy

Journal: Stem Cells and Cloning : Advances and Applications

Article Title: Characterization of neural stemness status through the neurogenesis process for bone marrow mesenchymal stem cells

doi: 10.2147/SCCAA.S94545

Figure Lengend Snippet: After 72 hours of culturing mouse MSCs in MEM +20% FBS, note that some of the cells became adherent and began to elongate as revealed under inverted microscope. Notes: ( A )10×; ( B )20×. Abbreviations: MSCs, mesenchymal stem cells; MEM, minimum essential medium; FBS, fetal bovine serum.

Article Snippet: There were four different formulas: 1) MEM media with 5% FBS alone, 2) MEM supplemented with 50 ng/mL epidermal growth factors (EGFs; United States Biological, MA, USA; E3374-11), 3) MEM supplemented with 50 ng/mL basic fibroblast growth factors (b-FGFs) (United States Biological; F4210-20M1), and 4) MEM with 50 ng/mL for each b-FGF and EGF.

Techniques: Inverted Microscopy