msd v-plex plus proinflammatory panel 1 mouse kit  (Meso Scale Diagnostics LLC)

Journal: Frontiers in Cardiovascular Medicine

Article Title: PPARβ/δ Is Required for Mesenchymal Stem Cell Cardioprotective Effects Independently of Their Anti-inflammatory Properties in Myocardial Ischemia-Reperfusion Injury

doi: 10.3389/fcvm.2021.681002

Figure Lengend Snippet: C57BL/6J mouse hearts were mounted on a Langendorff system and subjected to IR injury. (A) The ex vivo protocol included a 15 min-stabilization period, followed by 30 min of global ischemia achieved by stopping the flow in the aorta (no-flow). Reperfusion was achieved by restoring the Tyrode infusion during 60 min. For the SHAM condition, the heart was perfused throughout the protocol without any ischemic induction. At the end of the protocol, infarct size was measured using the TTC-staining method. Coronary effluents were collected at two time points during the ex vivo protocol: during stabilization to evaluate the “basal” level of cytokine release and at the end of the reperfusion phase to evaluate the “IR60” cytokine production after IR injury. (B) Scatter plots and bars (mean ± SD) were represented for infarct size (in % of LV) in IR ( n = 12) and SHAM ( n = 3) hearts. Representative pictures of TTC-stained LV slices were shown for each group. (C) Scatter plots with bars (mean ± SD) are presented for quantification of cytokines within coronary effluents collected before ischemia (Basal) and after 60 min of reperfusion after the IR protocol (IR 60 ) using the Meso Scale Discovery (MSD) V-Plex Plus Proinflammatory Panel 1 (mouse) kit. Statistical analysis was performed using the Mann-Whitney test. For CXCL1 (pg/ml), **** was noted for p < 0.0001, for IL-6 (pg/ml), **** was noted for p < 0.0001 and for TNFα (pg/ml), *** was noted for p = 0.0008.

Article Snippet: The different perfusates were stored at −80°C until the assay was performed using the Meso Scale Discovery (MSD) V-Plex Plus Proinflammatory Panel 1 (mouse) kit at the “Plateforme de Protéomique Clinique de Montpellier” according to the manufacturer's protocol.

Techniques: Ex Vivo, Staining, MANN-WHITNEY

Journal: Frontiers in Cardiovascular Medicine

Article Title: PPARβ/δ Is Required for Mesenchymal Stem Cell Cardioprotective Effects Independently of Their Anti-inflammatory Properties in Myocardial Ischemia-Reperfusion Injury

doi: 10.3389/fcvm.2021.681002

Figure Lengend Snippet: (A) Isolated hearts perfused ex vivo on the Langendorff system were submitted to the perfusion protocol similar to that described in . In the MSC group, reperfusion was achieved with a MSC Tyrode solution (5,000 cells/ml). For the PostC group, a postconditioning stimulus comprising three cycles of 1 min ischemia-1 min reperfusion was applied at the onset of reperfusion. In the control condition (IR), hearts were reperfused with Tyrode solution alone (control condition). Coronary effluents were collected at the end of the reperfusion phase to evaluate cytokine production after IR, PostC or MSC protocols. (B–D) Scatter plots with bars (mean ± SD) are presented for quantification of cytokines within coronary effluents collected after 60 min of reperfusion using the Meso Scale Discovery (MSD) V-Plex Plus Proinflammatory Panel 1 (mouse) kit. Statistical analysis was performed using the Kruskal-Wallis test followed by the Dunn's post test. (B) For CXCL1 (pg/mL), *** was noted for p = 0.0006 (PostC vs. IR), ** for p = 0.018 (MSC vs. IR) and ns for p > 0.999 (MSC vs. PostC). (C) For IL-6 (pg/ml), ** was noted for p = 0.0021 (PostC vs. IR), * for p = 0.0287 (MSC vs. IR) and ns for p > 0.999 (MSC vs. PostC) and (D) : for TNFα (pg/ml), ns was noted for p = 0.059.

Article Snippet: The different perfusates were stored at −80°C until the assay was performed using the Meso Scale Discovery (MSD) V-Plex Plus Proinflammatory Panel 1 (mouse) kit at the “Plateforme de Protéomique Clinique de Montpellier” according to the manufacturer's protocol.

Techniques: Isolation, Ex Vivo, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: PPARβ/δ Is Required for Mesenchymal Stem Cell Cardioprotective Effects Independently of Their Anti-inflammatory Properties in Myocardial Ischemia-Reperfusion Injury

doi: 10.3389/fcvm.2021.681002

Figure Lengend Snippet: (A) Isolated hearts perfused ex vivo on the Langendorff system were submitted to perfusion protocol similar to that described in . Reperfusion was achieved with a Tyrode solution alone in the IR group or with a Tyrode solution prepared at a concentration of 5,000 cells/ml with MSC (MSC group) and with KO MSC (KO MSC group). At the end of the protocol, infarct size analysis was performed on the isolated heart (B) and cytokine production was analyzed (C–E) using the Meso Scale Discovery (MSD) V-Plex Plus Proinflammatory Panel 1 (mouse) kit. Statistical analysis was performed using the Kruskal-Wallis test with the Dunn's post test. (B) Scatter plots and bars (mean ± SD) were represented for infarct size (in % of LV) in IR ( n = 12), MSC (5,000 cells/ml, n = 9), and KO MSC (5,000 cells/ml, n = 13). Statistical significance is noted ** for p = 0.001 (MSC vs IR), * for p = 0.029 (KO MSC vs. IR), and ns for p = 0.075 (KO MSC vs. MSC). (C–E) Scatter plots with bars (mean ± SD) are presented for quantification of TNFα, CXCL1, and IL-6 within coronary effluents collected at 60 min after the onset of reperfusion from untreated hearts (IR), hearts treated with wild-type MSC (MSC) and MSC deficient for PPARβ/δ (KO MSC) using the Meso Scale Discovery kit. Statistical analysis was performed using the Kruskal-Wallis test. (C) For CXCL1 (pg/ml), ** was noted for p = 0.0017 (MSC vs. IR), ns was noted for p = 0.34 (KO MSC vs. IR) and for p = 0.28 for (KO MSC vs. MSC); (D) For IL-6 (pg/ml), ** was noted for p = 0.0021 (MSC vs. IR), * for p = 0.028 (KO MSC vs. IR), and ns for p > 0.999 (KO MSC vs. MSC); (E) for TNFα (pg/ml), ns was noted for p = 0.60.

Article Snippet: The different perfusates were stored at −80°C until the assay was performed using the Meso Scale Discovery (MSD) V-Plex Plus Proinflammatory Panel 1 (mouse) kit at the “Plateforme de Protéomique Clinique de Montpellier” according to the manufacturer's protocol.

Techniques: Isolation, Ex Vivo, Concentration Assay