295 MSCs conditioned medium inhibits TNF-α induced human umbilical vein endothelial cells injury
Monday November 16, 2015 from 11:00 to 12:30
Room 111-112

Meimei Shi, People's Republic of China

West China Hospital, Sichuan University


MSCs conditioned medium inhibits TNF-α induced human umbilical vein endothelial cells injury

Meimei Shi1, Yujia Yuan1, Jingping Liu1, Younan Chen1, Bo Chen1, Lan Li1, Jie Zhang1, Ruixi Luo1, Yuanming Li1, Linzhao Li1, Yanrong Lu1, Jingqiu Cheng1.

1Laboratory of Transplant Engineering and Immunology, West China Hospital, Sichuan University, Chengdu, People's Republic of China

Transplant Engineering and Immunology, West China Hospital.

Objectives: Endothelial dysfunction caused by pro-inflammatory cytokines plays a crucial role in the pathogenesis of transplant rejection, thrombosis, diabetes and cardiovascular disease. Recent studies have shown that mesenchymal stem cells (MSCs) possess immune-modulatory, anti-apoptotic and anti-oxidant properties, which indicates their great advantages in clinical applications. In the present study, we investigate the impact of MSCs conditioned medium (MSCs-CM) on TNF-α induced human umbilical vein endothelial cells (HUVECs) injury.
Methods: Human umbilical cord MSCs were cultured in serum-free DMEM medium for 24 hours, MSCs-CM was prepared by concentrating the collected media for 10 folds via ultrafiltration. HUVECs were exposed to TNF-α (1ng/mL) with or without MSCs-CM for 72 hours. Cell viability was detected with CCK8 kit and cellular reactive oxygen species (ROS) were detected by flow cytometry after staining with DCFH-DA. The gene expression of inflammatory factors after 24 hours of TNF-α exposure was analyzed by real-time quantitative PCR. NF-κB activation was evaluated by nuclear translocation using immunofluorescence staining. In addition, the functions of HUVECs were evaluated by THP-1 cell adhesion and tube formation assay.
Results: The cell viability was declined to 72.4% of control group after the exposure to TNF-α, while MSCs-CM treatment restored the viability to 87.6%. In terms of oxidative stress, MSCs-CM reduced TNF-α induced ROS accumulation in HUVECs from 1.83 fold to 1.36 fold (P<0.01). Moreover, MSCs-CM treatment significantly inhibited the gene expression of VCAM-1, ICAM-1, MCP-1 and IL-6 that were up regulated by TNF-α. NF-κB was shown to be translocated into nuclear after TNF-α exposure, while MSCs-CM ameliorated this phenomenon. The nuclear/cytoplasmic (N/C) ratio was 0.45 vs.0.30 in TNF-α and MSCs-CM treatment group respectively. In addition, the number of adherent THP-1 cells was reduced by 52.5% in MSCs-CM group. Meanwhile, the tube formation capacity was improved by MSCs-CM, evidenced by increased tubular length (68% vs. 47%), branching point number (66% vs. 38%) and junctional areas (78% vs. 43%), the data was presented as the percentages compared to control group.
Conclusion: Our results indicate that MSCs-CM treatments are capable to preserve cell viability, attenuate cellular ROS accumulation, as well as inhibit the activation of NF-κB, consequently alleviating TNF-α induced HUVECs dysfunction. The study may provide a clue for considering MSCs conditioned medium as a potential cell-free strategy to ameliorate the inflammatory injury in endothelial cell.

Lectures by Meimei Shi

© 2018 Melbourne2015