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: Korean Journal of Neurotrauma

Article Title: Evaluation of Bone Marrow-derived Stem Cells and Adipose-derived Stem Cells Co-cultured on Human Nucleus Pulposus Cells: A Pilot Study

doi: 10.13004/kjnt.2020.16.e36

Figure Lengend Snippet: SA-β-gal: senescence associated-β-gal, NP: nucleus pulposus, NPC: nucleus pulposus cell, MSC: mesenchymal stem cell, ADMSC: adipose-derived mesenchymal stem cell, BDMSC: bone marrow-derived mesenchymal stem cell.

Article Snippet: BDMSC complete culture medium (ATCC ® PCS-500-030) plus one MSC growth kit (ATCC ® PCS-500-041) and ADMSC complete culture medium (ATCC ® PCS-500-030) plus one MSC growth kit (ATCC ® PCS-500-040) were used to cultivate both MSCs; this medium consists of 485 mL Human MSC Basal Medium, 35 mL Human MSC-Qualified FBS, 0.5 mL penicillin/streptomycin-amphotericin and 6 mL L-alanyl-L-glutamine.

Techniques: Derivative Assay

Journal: International Journal of Nanomedicine

Article Title: Drug Delivery to the Bone Microenvironment Mediated by Exosomes: An Axiom or Enigma

doi: 10.2147/IJN.S307843

Figure Lengend Snippet: Patents Related to the Diagnostic and Therapeutic Application of Exosomes Towards Bone Disorders

Article Snippet: Bone marrow-derived mesenchymal stem cells (BM-MSCs) and Adipose-derived stem cells (ASCs) , WO2019139762A1 , Zen-Bio, Inc. (US) , Exosome compositions and use thereof for joint disorders and diseases , Filed on 2018/Status pending.

Techniques: Diagnostic Assay, Biomarker Discovery, Derivative Assay, CRISPR, Isolation, Injection, Clinical Proteomics

Journal: The EMBO Journal

Article Title: Hypoxia promotes osteogenesis by facilitating acetyl‐CoA ‐mediated mitochondrial–nuclear communication

doi: 10.15252/embj.2022111239

Figure Lengend Snippet: A Schematic representation demonstrating the isolation protocol and the culture conditions of bone MSCs isolated from the back limbs of young (~3–5 months old) mice. After collection of the limbs, clean bones were cut into small pieces, which were then treated with collagenase for 1 h at 37°C. Cells and bone fragments were seeded in flasks and incubated for 10 days under 2% O 2 . On day 10, cell sorting was performed using flow cytometry and selecting the CD45 − /Ter‐119 − /Sca‐1 + /CD140a + mesenchymal stem cell population. The isolated population was then split into two groups: one was transferred back to hypoxia, whereas the other one was shifted to normoxia. Cells were cultured under these conditions for 7 days. B, C Representative images (B) and quantification (C) of Oil Red O staining of hypoxia‐ and normoxia‐cultured cells, 9 days after induction of adipogenesis. n = 5 biologically independent replicates. Results are shown as mean ± SEM and statistical significance was determined using a two‐sided unpaired t ‐test. D, E Representative images (D) and quantification (E) of Alizarin Red S staining of hypoxic, normoxic and reversed hypoxic (R_2% O 2 ) cells, 12 days after induction of osteogenesis. Cells were exposed to 21% O 2 for 7 days and then moved back to 2% O 2 , where osteogenesis was induced after 4 days. n = 3 for hypoxic cells and n = 5 biologically independent experiments for normoxic and reversed‐hypoxic cells, and merged results are shown in (E). Results are shown as mean ± SEM and statistical significance was determined with ordinary one‐way ANOVA, using Holm–Sidak's multiple‐comparisons test. F Principal component analysis (PCA) plot showing clustering of hypoxia‐ and normoxia‐cultured cells after RNA‐seq. n = 4 biologically independent replicates. G GO enrichment analysis for down‐regulated genes upon exposure to normoxia, as identified by RNA‐seq. Data information: Scale bars, 500 μm. Source data are available online for this figure.

Article Snippet: Commercial bone‐mesenchymal stem cells (MSCs) , Cyagen , MUBMX‐01001.

Techniques: Isolation, Incubation, FACS, Flow Cytometry, Cell Culture, Staining, RNA Sequencing Assay

Journal: The EMBO Journal

Article Title: Hypoxia promotes osteogenesis by facilitating acetyl‐CoA ‐mediated mitochondrial–nuclear communication

doi: 10.15252/embj.2022111239

Figure Lengend Snippet:

Article Snippet: Commercial bone‐mesenchymal stem cells (MSCs) , Cyagen , MUBMX‐01001.

Techniques: Transgenic Assay, Isolation, Recombinant, Generated, Software, Bicinchoninic Acid Protein Assay, RNA Extraction, Ligation, Mass Spectrometry, Liquid Chromatography, Microscopy

Journal: Experimental cell research

Article Title: High-content image informatics of the structural nuclear protein NuMA parses trajectories for stem/progenitor cell lineages and oncogenic transformation

doi: 10.1016/j.yexcr.2016.12.018

Figure Lengend Snippet: (A) Phase contrast images of MSCs, before (left) or after (right) the oncogenic NiSO4 treatment. (B) Proliferation of NiSO4-treated MSCs and controls measured using BrdU labeling after 3 days in culture. (C) Expression of the cell surface markers CD44, CD133, and Stro-1 quantified by flow cytometry in untreated and NiSO4-treated MSCs. (D) Expression of topoisomerase II-α, measured by immunostaining in MSCs and NiSO4-treated MSCs after 3 days in culture. (E) Telomerase (hTR) expression detected by FISH either 3 or 11 days after ending the NiSO4 treatment, as well as in untreated MSCs and in transformed MSCs (tMSCs). Fluorescence signal intensities are quantified in the bar graph and representative images are shown. (F) High-content analysis of nuclear descriptors derived from MSCs, NiSO4-treated MSCs (3 days after treatment), and tMSCs. Classification is visualized in 3D space after principal component analysis as well as with the parsing index. (G) Transformation indexes for MSC cells cultured on different biomaterial substrates (listed in the table) after NiSO4 exposure. The indexes represent hTR expression relative to untransformed MSCs (value 0) and tMSCs (value 1). hTR analysis was performed 11 days after ending the oncogenic treatment. (H) Highcontent imaging parsing indexes plotted against Transformation Indexes for MSCs cultured on different substrates as in (G). R, Pearson correlation coefficient. *, P < 0.01.

Article Snippet: Human mesenchymal stem cell culture and transformation MSCs were obtained from Lonza and cultured according to the supplier’s recommendations and reagents.

Techniques: Labeling, Expressing, Flow Cytometry, Immunostaining, Transformation Assay, Fluorescence, High Content Screening, Derivative Assay, Cell Culture, Imaging