Journal: Cells

Article Title: Interferon-Gamma Primed Human Clonal Mesenchymal Stromal Cell Sheets Exhibit Enhanced Immunosuppressive Function

doi: 10.3390/cells11233738

Figure Lengend Snippet: Duration of IFN-γ priming is directly related to enhanced hcBMSC sheet gene expression of immunomodulatory factors. Quantitative real-time PCR gene expression for immunomodulatory genes ( a ) human leukocyte antigen DR (HLA-DRA), ( b ) programmed death ligand-1 (PD-L1), ( c ) indoleamine 2,3-dioxygenase (IDO-1), ( d ) interleukin 10 (IL-10), and ( e ) prostaglandin E synthase 2 (PTGES2). All gene expression is normalized to GAPDH, fold change is relative to non-primed control hBMSC sheet sample. Error bars represent means ± SE ( n = 6) (n.s. p ≥ 0.05, ** p < 0.01, *** p < 0.001) Statistical analysis p values are compared to non-IFN-γ hcBMSC sheets (non-IFN-γ, control).

Article Snippet: The concentration of soluble IL-10, IDO, PGE2, and PD-L1 per sample was quantified using human IL-10 DuoSet ELISA kit (R&D Systems, Minneapolis, MN, USA), human indoleamine 2,3-dioxygenase/IDO DuoSet ELISA kit (R&D Systems, Minneapolis, MN, USA), prostaglandin E2 Parameter Assay Kit (R&D Systems, Minneapolis, MN, USA), and human PD-L1 DuoSet ELISA Kit (R&D Systems, Minneapolis, MN, USA), respectively.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Control

Journal: Journal of Translational Medicine

Article Title: Manufacturing mesenchymal stromal cells in a microcarrier-microbioreactor platform can enhance cell yield and quality attributes: case study for acute respiratory distress syndrome

doi: 10.1186/s12967-024-05373-7

Figure Lengend Snippet: Protein Expression in gelatin microcarrier-microbioreactor and T25 Flask. The term ‘therapeutic’ in green font signals an anticipated correlation with improved therapeutic efficacy in ARDS, as hypothesized based on review of existing literature. The term ‘Pro-inflammatory’ in red font signals an anticipated correlation with an anti-therapeutic effect in ARDS, as hypothesized by review of existing literature. MSC gene expression was evaluated at the protein level for a subset of genes from Fig. to confirm protein translation and evaluate the absolute concentration of effector molecule present. Protein-level data corroborated some – though not all – of the trends reported from mRNA-level data. Protein concentration was evaluated from MSC secretome by harvesting the conditioned media from culture days 5–7 and utilizing colorimetric ELISA for each analyte. Analytes were assessed at two dilutions, each in triplicate wells, for each condition. Protein concentration was normalized to cell count. Conditioned media was vortexed with 0.02% Tween20 to rupture liposomes prior to starting the ELISA protocol. Remaining steps followed the manufacturers’ protocols. Significance bars indicate results of unpaired two-tailed t-tests. *, p < 0.05. GMC-MB: Gelatin microcarrier-microbioreactor

Article Snippet: Colorimetric sandwich ELISA kits (R&D Systems, DY286-05, DY6030B-05; Thermo Scientific, 88-7176-86, BMS2034MST; Fisher Scientific, CHC1323) were used to quantify secretome protein expression.

Techniques: Expressing, Concentration Assay, Protein Concentration, Enzyme-linked Immunosorbent Assay, Cell Counting, Liposomes, Two Tailed Test