Journal: Retrovirology

Article Title: Extracellular vesicles from HTLV-1 infected cells modulate target cells and viral spread

doi: 10.1186/s12977-021-00550-8

Figure Lengend Snippet: Functional Effects of HTLV-1 EVs on Angiogenesis and Inflammation. a An angiogenesis assay in technical and biological triplicate was used to determine the effect of distinct HTLV-1 EVs (2 k, 10 k, and 100 k) on tubular formation in a co-culture of mesenchymal stem cells (MSCs) and aortic endothelial cells (AEC). EV-treated cells (1:2000 recipient cell to EV ratio). Positive control cells received complete, undiluted medium. Additional dose of EV treatment was given to the cells at day 4. Representative images were taken at days 3 and 6 showing tubular formation in response to the indicated treatment. b The image processing software WIMASIS was used to calculate the percentage of area covered by tubules (n = 3) on day 3 and day 6. c RT-qPCR results showing env RNA copy numbers of mesenchymal stem cells treated with CEM EVs (control) and different populations of HTLV-1 EVs: 2 k (left panel), 10 k (middle panel), and 100 k (right panel). A set of dashed black vertical lines (---) were used to indicate baseline env RNA copy numbers. A set of dashed red vertical lines ( ) were used to indicate the levels of starting material, suggestive of the minimum env RNA copy numbers necessary for EVs to increase vial spread in MSc. d Western blot analysis for core histones (H3, H2A, H2B, and H4), linker histone (H1) and actin in HUT102 EVs (2 k, 10 k, and 100 k). e GAPDH DNA levels (representative of nucleosomes) in 2 k, 10 k, and 100 k HUT102 EVs treated with proteinase K and DNase/RNase were evaluated by was quantitated by q-PCR. A two-tailed student t-test was used to evaluate statistical significance with “**” for p-values ≤ 0.01, indicating the level of significance relative to untreated (Control) samples

Article Snippet: Bone marrow derived Mesenchymal Stem Cells (MSCs) were cultured in Mesenchymal Stem Cell Basal Medium (ATCC PCS-500–030TM) supplemented with Mesenchymal Stem Cell Growth Kit for Bone Marrow-derived MSCs (ATCC PCS-500-041TM).

Techniques: Functional Assay, Angiogenesis Assay, Co-Culture Assay, Positive Control, Software, Quantitative RT-PCR, Control, Western Blot, Two Tailed Test

Journal: Cells

Article Title: Combined TGF-β3 and FGF-2 Stimulation Enhances Chondrogenic Potential of Ovine Bone Marrow-Derived MSCs

doi: 10.3390/cells14131013

Figure Lengend Snippet: Alcian blue staining of ovine BM-MSCs in 2D culture in ( a ) chondrogenic differentiation medium ( b ) with FGF-2, ( c ) TGF-β3, ( d ) FGF-2, and TGF-β3. ( e ) Effect of TGF-β3 and FGF-2 cytokines on the intensity of Alcian blue staining in 2D culture for 21 days of culture. Three independent experiments in triplicates were performed. * p < 0.05. Ovine BM-MSCs treated with FGF-2 and TGF-β3 exhibit the highest chondrogenic differentiation potential.

Article Snippet: After an overnight attachment, the medium was replaced with the chondrogenic differentiation medium (PromoCell, Heidelberg, Germany, cat. no. C-28016) either supplemented or not with FGF-2 or TGF-β3 or both.

Techniques: Staining

Journal: Cells

Article Title: Combined TGF-β3 and FGF-2 Stimulation Enhances Chondrogenic Potential of Ovine Bone Marrow-Derived MSCs

doi: 10.3390/cells14131013

Figure Lengend Snippet: Alcian blue staining of ovine (BM-MSCs) in 3D culture in ( a ) chondrogenic differentiation medium ( b ) supplemented with FGF-2 ( c ) TGF-β3, ( d ) FGF-2, and TGF-β3. ( e ) Intensity of Alcian blue staining of sheep BM-MSCs in 3D culture for 21 days. Three experiments in triplicates were performed. * p ˂ 0.05, ** p ˂ 0.005, and *** p ˂ 0.0001. Treatment of sheep BM-MSCs with both FGF-2 and TGF-β3 enhances 3D chondrogenic differentiation.

Article Snippet: After an overnight attachment, the medium was replaced with the chondrogenic differentiation medium (PromoCell, Heidelberg, Germany, cat. no. C-28016) either supplemented or not with FGF-2 or TGF-β3 or both.

Techniques: Staining

Journal: Cells

Article Title: Combined TGF-β3 and FGF-2 Stimulation Enhances Chondrogenic Potential of Ovine Bone Marrow-Derived MSCs

doi: 10.3390/cells14131013

Figure Lengend Snippet: Real-time PCR analysis for markers of chondrogenic differentiation of sheep BM-MSCs treated with TGF-β3 and/or FGF-2 for 7, 14, and 21 days. Chad , Comp , and Sox 5 ( a – c ) characterize the early stage of chondrogenesis; gene expression of Agg , Col IX , Sox 9 , and Fmod ( d – g ) characterize the late stage of chondrogenic differentiation. * p ˂ 0.05, ** p ˂ 0.005, and *** p ˂ 0.0001. Three experiments with two biological replicates were conducted. Treatment of ovine BM-MSCs with the cytokines FGF-2 and TGF-β3 increase the expression of both early and late chondrogenic marker genes.

Article Snippet: After an overnight attachment, the medium was replaced with the chondrogenic differentiation medium (PromoCell, Heidelberg, Germany, cat. no. C-28016) either supplemented or not with FGF-2 or TGF-β3 or both.

Techniques: Real-time Polymerase Chain Reaction, Gene Expression, Expressing, Marker