Journal: Cell Transplantation

Article Title: Comparing Scaffold-Free and Fibrin-Based Adipose-Derived Stromal Cell Constructs for Adipose Tissue Engineering: An In Vitro and in Vivo Study

doi: 10.3727/096368912x653129

Figure Lengend Snippet: Figure 4. ASCs in fibrin-based constructs secrete higher levels of the angiogenic factors VEGF and HGF than ASC in spheroids. The secretion of the angiogenic factors fibroblast growth factor 2 (FGF2), angiopoietin (ANG2), platelet-derived growth factor (PDGF), tissue inhibitors of matrix metalloproteinases 2 (TIMP2), TIMP1, hepatocyte growth factor (HGF), and vascular endothe- lial growth factor (VEGF) by ASCs in spheroids and fibrin-based constructs over 48 h was measured by multiplex chemiluminescent ELISA at day 7. Unconditioned adipogenic medium was used as control condition. Values are presented as median (interquartile range); n = 5, 3 ASC donors. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 (fibrin-based construct conditioned medium vs. spheroid conditioned medium and control).

Article Snippet: A human cytokine inflammation multiplex chemiluminescent ELISA kit (Cat. No. 110451HU, Quansys Biosciences, West Logan, UT, USA) and a human cytokine inflammation multiplex chemiluminescent ELISA kit (cat. no.150251HU; Quansys Biosciences) were used to assay conditioned and unconditioned medium for the release of the following inflammatory cytokines and angiogenic factors: interleukin-1a (IL-1a), IL-1b, IL-2, IL-4, IL-6, IL-8, IL-10, tumor necrosis factor-a (TNF-a), interferon-g (IFN-g), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), angiopoietin 2 (ANG2), fibroblast growth factor 2 (FGF2), hepatocyte growth factor (HGF), and the tissue inhibitors of metalloproteinases (TIMP): TIMP1, and TIMP2.

Techniques: Construct, Derivative Assay, Multiplex Assay, Chemiluminescent ELISA, Control

Journal: Cell Transplantation

Article Title: Comparing Scaffold-Free and Fibrin-Based Adipose-Derived Stromal Cell Constructs for Adipose Tissue Engineering: An In Vitro and in Vivo Study

doi: 10.3727/096368912x653129

Figure Lengend Snippet: Figure 6. ASCs in fibrin-based constructs display increased adipogenic differentiation when compared to ASCs in spheroids. (A) ASCs in spheroids and fibrin-based constructs were cultured for 7 days in vitro in adipogenic medium. Cross-sections were stained with Oil Red O (showing intracellular lipid) and counterstained with haematoxylin. (A.1) Spheroid. (A.2) Fibrin-based construct. Scale bars: 100 µm. (B) Q-PCR was used to measure the expression levels of the adipocyte-specific markers peroxisome proliferator-activated receptor g (PPARG), perilipin 1 (PLIN 1), fatty acid binding protein 4 (FABP4), and leptin (LEP). Expression levels are relative to b-2- microglobulin-positive control housekeeping gene (dCt). Values are presented as medians (interquartile range). For each of the three ASC donors, nine spheroids, nine fibrin-based constructs, and 0.5 ´ 106 undifferentiated ASCs (=control) were prepared and assayed. *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001 (relative expression in fibrin-based constructs compared to spheroids and control). (C) The secre- tion of leptin by ASCs in conditioned medium of spheroids and fibrin-based constructs over 48 h was measured by sandwich ELISA at day 7. Unconditioned adipogenic medium was used as control condition. Values are presented as median (interquartile range); n = 9, 3 ASC donors. **p ≤ 0.01, ***p ≤ 0.001 (fibrin-based construct conditioned medium vs. spheroid conditioned medium and control).

Article Snippet: A human cytokine inflammation multiplex chemiluminescent ELISA kit (Cat. No. 110451HU, Quansys Biosciences, West Logan, UT, USA) and a human cytokine inflammation multiplex chemiluminescent ELISA kit (cat. no.150251HU; Quansys Biosciences) were used to assay conditioned and unconditioned medium for the release of the following inflammatory cytokines and angiogenic factors: interleukin-1a (IL-1a), IL-1b, IL-2, IL-4, IL-6, IL-8, IL-10, tumor necrosis factor-a (TNF-a), interferon-g (IFN-g), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), angiopoietin 2 (ANG2), fibroblast growth factor 2 (FGF2), hepatocyte growth factor (HGF), and the tissue inhibitors of metalloproteinases (TIMP): TIMP1, and TIMP2.

Techniques: Construct, Cell Culture, In Vitro, Staining, Expressing, Binding Assay, Positive Control, Control, Sandwich ELISA

Journal: Frontiers in Aging Neuroscience

Article Title: Discovery of small molecule mechanistic target of rapamycin inhibitors as anti-aging and anti-cancer therapeutics

doi: 10.3389/fnagi.2022.1048260

Figure Lengend Snippet: Secretion of pro-inflammatory cytokines by senescent cells after exposure to compound 3 and torkinib. Determination of 11 pro-inflammatory cytokines IL-1α, IL-1β, IL-6, IL-8, IL-10, IL-12p70, IL-27, MCP-1, IFNγ, TNFα, and IP-10, and IP-10 in docetaxel-(DIS-BJ), ionizing-radiation- (IR-RPE-1), and temozolomide-induced (TMZ-U87) senescent cells exposed to compound 3 and torkinib (50 μM) for 24 h. Conditioned media were collected 24 h after treatment and analyzed for the level of cytokines by FACS analysis. The experiment was done in triplicate (TMZ-U87 in duplicate). Data were normalized to untreated cells as fold induction to control (set as 1, red line) and plotted as mean ± SEM. ND, not detected. Two-tailed Student’s t -test, * p < 0.05.

Article Snippet: To determine the secretion of selected inflammatory cytokines into culture media, cell cultures were exposed to tested compounds (50 μM) for 24 h. Then the culture medium was removed, cells were washed three times with 1 mL PBS, and a fresh medium without FBS was added for 24 h. Detection of cytokines in conditioned culture media was done by Human Inflammation 11-Plex Panel: IFNγ, IL-1α, IL-1β, IL-6, IL-8, IL-10, IL-12p70, IL-27, IP-10, MCP-1, and TNFα (AimPlex Biosciences, Inc.) using a BD FACSVerseTM flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) according to manufacturer’s protocol.

Techniques: Two Tailed Test