Journal: Molecular Medicine Reports

Article Title: A combined antitumor strategy of separately transduced mesenchymal stem cells with soluble TRAIL and IFNβ produces a synergistic activity in the reduction of lymphoma and mice survival enlargement

doi: 10.3892/mmr.2022.12722

Figure Lengend Snippet: BM-MSCs characterization. (A) BM-MSCs were adherent to the culture flask and presented elongated morphology (scale bar, 100 µm). They also expressed high mesenchymal markers, such as (B) CD90 and (C) CD105 (scale bar, 100 µm). In addition, they could differentiate to chondrogenic by (D) Alcian blue staining and calcified areas generated by osteogenic cells with (E) Von Kossa stain. BM-MSCs, bone marrow mesenchymal stem cells.

Article Snippet: MSC multipotency was examined by inducing the cells to differentiate to osteoblasts or chondroblasts, using the appropriate reagents in the Mouse Mesenchymal Stem Cell Functional Identification kit (cat. no. SC010; R&D Systems, Inc.) and following the manufacturer's instructions.

Techniques: Staining, Generated

Journal: Molecular Medicine Reports

Article Title: A combined antitumor strategy of separately transduced mesenchymal stem cells with soluble TRAIL and IFNβ produces a synergistic activity in the reduction of lymphoma and mice survival enlargement

doi: 10.3892/mmr.2022.12722

Figure Lengend Snippet: Lentiviral transduction of BM-MSC. (A-C) Lentiviral vectors were designed to contain (A) sTRAIL, (B) flTRAIL and (C) IFNβ. (D) Representative fluorescent images of BM-MSCs transduced with lentiviral vectors; nucleus stained with DAPI (blue) and transduced BM-MSCs expressing GFP protein (green signal). (E and F) Western blot analysis confirming protein expression of (E) flTRAIL and sTRAIL (F) and IFNβ in untransfected MSCs and MSCs transduced with a lentivirus that does not contain any of the genes of interest (empty lentiviral vector pLV[Exp]-EGFP:T2A:Puro-EF1A> mCherry, Vector Builder). MSCs express each of the genes flTRAIL, sTRAIL and IFN and as positive control the recombinant protein. Actin was used as a loading control. BM-MSCs, bone marrow mesenchymal stem cells; sTRAIL, TRAIL soluble; flTRAIL, TRAIL full length; IFNβ, interferon β.

Article Snippet: MSC multipotency was examined by inducing the cells to differentiate to osteoblasts or chondroblasts, using the appropriate reagents in the Mouse Mesenchymal Stem Cell Functional Identification kit (cat. no. SC010; R&D Systems, Inc.) and following the manufacturer's instructions.

Techniques: Transduction, Staining, Expressing, Western Blot, Plasmid Preparation, Positive Control, Recombinant, Control

Journal: Molecular Medicine Reports

Article Title: A combined antitumor strategy of separately transduced mesenchymal stem cells with soluble TRAIL and IFNβ produces a synergistic activity in the reduction of lymphoma and mice survival enlargement

doi: 10.3892/mmr.2022.12722

Figure Lengend Snippet: Kaplan-Meier curves. (A) First model with all treatments. (B) Second model with sTRAIL and IFNβ treatment and MSC naïve. *P<0.05 and **P<0.001 compared with the untreated group. sTRAIL, TRAIL soluble; IFNβ, interferon β; MSC, mesenchymal stem cell.

Article Snippet: MSC multipotency was examined by inducing the cells to differentiate to osteoblasts or chondroblasts, using the appropriate reagents in the Mouse Mesenchymal Stem Cell Functional Identification kit (cat. no. SC010; R&D Systems, Inc.) and following the manufacturer's instructions.

Techniques:

Journal: eLife

Article Title: Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

doi: 10.7554/eLife.47970

Figure Lengend Snippet: ( A ) Bar graph shows expression profile of MYH isoforms in hiPSC-1-derived myotubes. Data are shown as mean ± S.E.M.; n = 3, ***p<0.001. ( B ) Bar graph shows the ratio of % MyHC-stained area to % DAPI area in myotubes resulting from treatment with five candidates identified by the small molecule screening. Data show significant increase (***p<0.001) compared to DMSO in all three PS cell lines analyzed (hESC-1, hiPSC-1 and hiPSC-2). Data from three independent replicates are shown, normalized to DMSO, as mean ± S.E.M. ( C ) Bar graph shows the ratio of % MyHC-stained area to % DAPI area in iPS cell-derived myotubes that had been differentiated in the presence of all candidates combined, or with individual candidates excluded from the overall combination. Data from three independent replicates are shown normalized to DMSO. Values are shown as mean ± S.E.M. ***p<0.001. ( D ) Representative images show immunostaining for MyHC (in red) in hiPSC-1 myotubes differentiated with combinatory treatments of small molecules or DMSO. DAPI stains nuclei (in blue). Scale bar is 100 μm. ( E ) Bar graph shows fusion index analysis of myotubes that were differentiated with small molecule combinations or DMSO. Data are shown as mean of three independent replicates ± S.E.M. ***p<0.001. ( F ) Stacked bar graph shows the frequency of number of nuclei per myotube upon differentiation with combinatory treatments or DMSO. Data are shown as mean of three independent replicates ± S.E.M. Statistical analysis compares each combination to DMSO. *p<0.05 **p<0.01 ***p<0.001. 10.7554/eLife.47970.006 Figure 1—source data 1. Tocriscreen Stem Cell Toolbox compounds tested during myogenic terminal differentiation of PS cell lines.

Article Snippet: To determine whether small molecule compounds may enhance the maturation of PS cell-derived myotubes, we performed a small molecule library screening using the Tocriscreen stem cell toolbox kit (Tocris).

Techniques: Expressing, Derivative Assay, Staining, Immunostaining

Journal: eLife

Article Title: Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

doi: 10.7554/eLife.47970

Figure Lengend Snippet: ( A ) Schematic representation of the small molecule screening procedure. Each well from a 96-well plate contained an individual compound from the Tocriscreen Stem cell toolbox added in the differentiation medium. ( B ) Representative images of MyHC (red) immunostaining in myotubes differentiated with selected candidates upon small molecule screening ( A ). DAPI stains blue. Scale bar is 100 μm. ( C ) Bar graph shows ratio of % MyHC-stained area to % DAPI area from hiPSC-1 myotubes differentiated with compounds candidates at 5, 10 or 20 μm relative to DMSO. Data are shown as mean of three independent replicates ± S.E.M. Statistical analyses showed no significant differences among concentrations for each compound. ( D ) Bar graphs show gene expression analysis of MYOG and MYH isoforms relative to GAPDH of hiPSC-1 myotubes differentiated with compound candidates. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01 ***p<0.001.

Article Snippet: To determine whether small molecule compounds may enhance the maturation of PS cell-derived myotubes, we performed a small molecule library screening using the Tocriscreen stem cell toolbox kit (Tocris).

Techniques: Immunostaining, Staining, Expressing

Journal: eLife

Article Title: Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

doi: 10.7554/eLife.47970

Figure Lengend Snippet:

Article Snippet: To determine whether small molecule compounds may enhance the maturation of PS cell-derived myotubes, we performed a small molecule library screening using the Tocriscreen stem cell toolbox kit (Tocris).

Techniques: Control, Recombinant, Imaging, Proliferation Assay, Sequencing

Journal: Cell Death & Disease

Article Title: Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment

doi: 10.1038/cddis.2013.88

Figure Lengend Snippet: Characterization of HS-181 cell line after treatment with zebularine. ( a ) RT-PCR of cardiac markers after treatment with zebularine. Myh7, Myh6, Actc, cTnI and Serca2 present more expression after treatment. ( b ) Immunostaining of treated cells to detect cardiac-specific proteins. Scale bars; 50 μ m. ( c ) Blots of Proteome Profiler Array and the resulting quantification histograms demonstrating inhibition of pluripotency marker expression and ( d ) increased levels of mesodermic proteins after zebularine treatment (black arrows)

Article Snippet: Protein expression profiles were assayed using the specific human pluripotent Stem Cell array kit ‘Proteome Profiler Array' (R&D Systems Europe, Abingdon, UK; ARY010) following the manufacturer's instructions.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Immunostaining, Inhibition, Marker

Journal: BMC Cancer

Article Title: Tyrosine kinase inhibitor SU11274 increased tumorigenicity and enriched for melanoma-initiating cells by bioenergetic modulation

doi: 10.1186/s12885-016-2341-y

Figure Lengend Snippet: Bioenergetic modulation counteracts the protumorigenic action of the SU11274. a – b Protein lysates were prepared from untreated controls and Rel3 cells treated with 1 μM SU11274 in spheroid conditions for 7 days. Pluripotent stem cell array has shown expression of all pluripotency markers in Rel3 cells expanded as spheroids. SU11274 treatment further increased level of proteins associated with pluripotency in SU11274-treated cells correlating with their higher tumor initiating properties. Phosphokinase assay screening has shown several active intracellular signaling cascades. Phosphorylated forms of p53 (S392 and S46) were detected in the SU11274-treated cells (lower panel B), which correlates with SU11274 mediated inhibition of cell proliferation. RSK1/2/3 (S380) phosphorylation was increased in SU11274-treated cells. We detected phosphorylation of the following target kinases: ERK1/2 (T202/Y204, T185/Y187), P-RAS40 (T246), Akt 1/2/3 (S473), p38 alpha (T180/Y182), AMPK alpha1 (T183), CREB (S133), GSK-3 alpha/beta(S21/S9), WNK-1 (T60) and HSP60 in both treated and untreated cells

Article Snippet: Proteome profile of the melanosphere cells cultured for 7 days in the presence of 1 μM SU11274 was evaluated by the Proteome ProfilerTM Human Phospho-Kinase Antibody Array and the Human Pluripotent Stem Cell Antibody Array (R&D SystemsTM Inc., Minneapolis MN).

Techniques: Expressing, Inhibition, Phospho-proteomics