Journal: Journal of Tissue Engineering

Article Title: Therapeutic potential of adipose-derived mesenchymal stem cell exosomes in tissue-engineered bladders

doi: 10.1177/20417314211001545

Figure Lengend Snippet: Identification of Ad-MSCs (a–c) and AMEs (d–f): (a) characterization of the fourth generation of Ad-MSCs was performed by flow cytometry, and the Ad-MSC histogram revealed that after purification, the cells mostly expressed mesenchymal cell markers. Differentiation assays showed that Ad-MSCs could differentiate into adipocytes (b) and osteocytes (c), which were stained with Oil Red O and Von Kossa, respectively. Scale bar = 100 μm. (d) TEM of exosomes isolated from MSCM-cultured Ad-MSCs for 3 days with 10% exosome-depleted FBS. Scale bars = 100 nm. (e) NTA found that most of these vesicles ranged in diameter from 30 to 150 nm. (f) Western blotting was performed with Ad-MSCs (MSCs) and AMEs (Exos). The expression of CD9, CD63, TSG101, and calnexin was detected. Ad-MSCs: adipose-derived mesenchymal stem cells; AMEs: adipose-derived mesenchymal stem cell exosomes; NTA: nanoparticle tracking analysis; TEM: transmission electron microscopy.

Article Snippet: The medium was changed to mesenchymal stem cell adipogenic differentiation medium (MADM, ScienCell) and mesenchymal stem cell osteogenic differentiation medium (MODM, ScienCell) when the cells reached 70–80% confluency.

Techniques: Flow Cytometry, Purification, Staining, Isolation, Cell Culture, Western Blot, Expressing, Derivative Assay, Transmission Assay, Electron Microscopy

Journal: RMD Open

Article Title: Dual neutralisation of IL-17F and IL-17A with bimekizumab blocks inflammation-driven osteogenic differentiation of human periosteal cells

doi: 10.1136/rmdopen-2020-001306

Figure Lengend Snippet: IL-17 is intimately involved with osteogenic differentiation of hPDCs. (A) Expression of SOSDC1, RUNX2, IL-17RA and IL-17RC in hPDCs cultured in GM or GFC. Results are expressed as the mean fold change in expression compared to day 0 GM±SEM (n=3). (B) Representative phase-contrast images of hPDCs during 3, 5, 7 and 9 days of culture under GM or GFC conditions showing bone nodule formation (dark regions from day 7) in GFC, with corresponding day 9 alizarin red–stained monolayer images. (C) Expression of RUNX2 and BMP2 cultured under DM or supplemented with 50 ng/mL rhIL-17A or 50 ng/mL IL-17F for 9 days. Results are expressed as the mean fold change in expression compared to day 0 GM±SEM (D) Quantification of matrix mineralisation in hPDCs cultured in DM or supplemented with 50 ng/mL IL-17A or 50 ng/mL IL-17F for 9 days. Representative images of matrix mineralisation stained with alizarin red are presented below the quantification graph. Results are expressed as the mean fold change in expression compared to day 0 GM±SEM. ***p<0.001; **p<0.01; *p<0.05; comparisons between each condition by one-way ANOVA with Fisher’s LSD post hoc test (n=3). ANOVA, analysis of variance; DM, differentiation medium; GM, growth medium; GFC, growth factor cocktail; hPDCs, human periosteum-derived cells; IL, interleukin.

Article Snippet: A biomimetic human periosteum-derived cell (hPDC) model of osteogenic differentiation was used in combination with recombinant IL-17 cytokines, T-cell supernatants or serum from patients with AS.

Techniques: Expressing, Cell Culture, Staining, Derivative Assay

Journal: RMD Open

Article Title: Dual neutralisation of IL-17F and IL-17A with bimekizumab blocks inflammation-driven osteogenic differentiation of human periosteal cells

doi: 10.1136/rmdopen-2020-001306

Figure Lengend Snippet: Bimekizumab suppresses γδ T cell and Th17 supernatant-mediated osteogenic differentiation of hPDCs. Heat maps summarising inhibition of osteogenic marker expression in hPDCs stimulated with DM or DM supplemented with (A) γδ T cell supernatant or (B) Th17 cell supernatant (± preincubation with 10 µg/mL anti-IL-17A; anti-IL-17F or bimekizumab) for 9 days. Results are expressed as the Log 2 mean fold change in expression compared to growth media control. Quantification of matrix mineralisation after 9 days incubation in (C) γδ T cell supernatant or (D) Th17 cell supernatant (± preincubation with 10 µg/mL anti-IL-17A; anti-IL-17F or bimekizumab). Representative images of matrix mineralisation stained with alizarin red are presented below the quantification graphs. Results are expressed as the mean fold change in expression compared to GM±SEM. ***p<0.001; **p<0.01; *p<0.05; comparisons between each condition by one-way ANOVA with Fisher’s LSD post hoc test (n=3). (E) MSD-multi-array v-plex pro-inflammatory panel expression of IFNγ, TNFα, IL-1β, IL-8, IL-10, IL-17A, IL-17F cytokines from Th17 and γδT cell supernatants. Results are expressed as mean pg/mL ± SEM. ***p<0.001; **p<0.01; comparisons between Th17 and γδT cell supernatants by two-tailed Student’s t-test for each cytokine tested. (F) Expression of DKK1 in hPDCs stimulated with DM or DM supplemented with either γδ T cell or Th17 cell supernatant (± preincubation with 10 µg/mL anti-IL-17A; anti-IL-17F or bimekizumab) for 9 days. Results are expressed as the mean fold change in expression compared to day 0 GM ± SEM. ***p<0.001; **p<0.01; *p<0.05; comparisons between each condition by one-way ANOVA with Fisher’s LSD post hoc test (n=3). ANOVA, analysis of variance; DM, differentiation medium; GM, growth medium; hPDCs, human periosteum-derived cells; IL, interleukin; LSD, least significant difference; MSD, Meso Scale Discovery; Th17, T helper 17; TNF, tumor necrosis factor.

Article Snippet: A biomimetic human periosteum-derived cell (hPDC) model of osteogenic differentiation was used in combination with recombinant IL-17 cytokines, T-cell supernatants or serum from patients with AS.

Techniques: Inhibition, Marker, Expressing, Control, Incubation, Staining, Two Tailed Test, Derivative Assay

Journal: iScience

Article Title: Temporal transcriptome features identify early skeletal commitment during human epiphysis development at single-cell resolution

doi: 10.1016/j.isci.2023.107200

Figure Lengend Snippet:

Article Snippet: Mesenchymal stem cell osteogenic differentiation kit , iCell Bioscience Inc , Cat#iCEll-MSCYD-002.

Techniques: Staining, RNA Extraction, Reverse Transcription, Derivative Assay, Software, Imaging