Journal: Molecular Therapy. Nucleic Acids

Article Title: Ratio of miRNA-29 to miRNA-199 expression coordinates mesenchymal stem cell repair of bleomycin-induced pulmonary injury

doi: 10.1016/j.omtn.2025.102461

Figure Lengend Snippet: Bleomycin-induced lung injury, which is reduced after adipose-derived mesenchymal stem cell instillation (A) Representative μCT transverse and coronal lung sections acquired from aged (22-month-old) male C57BL/6 mice at baseline (left) and 7 days following intratracheal bleomycin (BLM, 2.0 U/kg) administration (right) demonstrating increased lung density and loss of airspaces. (B) Saline treatment did not result in evidence of lung injury on μCT scan at baseline (left) or 7 days post-instillation (right). Histological sections of lung tissue collected at day 21 post-BLM were stained with Masson’s trichrome as described in . (C and D) Representative photomicrographs (20× and 40× magnifications) of lung sections from saline-treated control mice (C) and BLM-treated mice (D). (E) Infusion of adipose-derived mesenchymal stem cells (ASCs) 12 days post-BLM instillation resulted in reduced severity of pulmonary fibrosis (PF). (F) Degree of PF on histological sections was measured by semi-quantitative Ashcroft score as described in . BLM-induced lung injury resulted in increased Ashcroft score compared to saline controls. Infusion with ASCs 12 days post-BLM injury resulted in decreased Ashcroft score. (G) Intratracheal BLM instillation increased lung collagen content as measured by hydroxyproline assays as described in . Mice treated with ASCs on day 12 post-BLM had decreased lung collagen content compared to BLM-only controls. Each data point represents an individual biological replicate (mouse); n = 6–10 mice/group. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. (H) Ratio of pAKT to AKT protein expression in lung tissue of mice was quantified by western blot analysis at day 21 post-BLM sacrifice. Aged C57BL/6 mice treated with intratracheal BLM demonstrated increased pAKT/AKT protein expression compared to saline-treated controls. Lungs from mice treated with intravenous infusion of ASCs 12 days post-BLM-induced injury demonstrated decreased expression of pAKT/AKT compared to BLM-only group. Inset shows a representative western blot and β-actin loading control. Data are graphed as individual biological replicates ( n = 6–8 mice/group); ∗ p < 0.05.

Article Snippet: For mesenchymal differentiation potential, the Mouse Mesenchymal Stem Cell Functional Identification Kit (R&D Systems, Minneapolis, MN) was used according to the manufacturer’s instructions.

Techniques: Derivative Assay, Saline, Staining, Control, Expressing, Western Blot

Journal: Scientific reports

Article Title: Small extracellular vesicles convey the stress-induced adaptive responses of melanoma cells.

doi: 10.1038/s41598-019-51778-6

Figure Lengend Snippet: Figure 1. Schematic illustration of the experimental workflow in six steps. B16F1 mouse melanoma cell- derived sEVs were isolated and characterised by Western Blot (WB), atomic force microscopy (AFM) and dynamic light scattering (DLS). Then B16F1 cultures were treated in five different ways, and 72 h supernatants were harvested for sEV isolation. Vesicle samples were then analysed by nanoparticle tracking analysis (NTA) to determine the number of released sEVs, sequencing and LC-MS/MS to describe their miRNome and proteome. Ingenuity Pathway Analysis (IPA) was used to analyse data and predict the functional differences between sEV groups. This in silico predictions were tested in vitro on mesenchymal stem cell (MSC) and melanoma cell cultures and MSC-B16F1 3D co-cultures as well using Ki-67-specific immunocytochemistry, Cell-Clock cell cycle assay, wound healing assay, and 3D hanging drop technology. Abbreviation: n.ctrl-negative control. Figure was created with BioRender.com.

Article Snippet: Purity of MSC cultures was checked by flow cytometry using the Mouse Multipotent Mesenchymal Stromal Cell Marker Antibody Panel (R&D Systems) according to the manufacturer’s instructions and a FACSCalibur instrument coupled with CellQuest Pro 6.0 (BD Biosciences).

Techniques: Derivative Assay, Isolation, Western Blot, Microscopy, Sequencing, Liquid Chromatography with Mass Spectroscopy, Functional Assay, In Silico, In Vitro, Immunocytochemistry, Cell Cycle Assay, Wound Healing Assay, Negative Control