689: Mesenchymal stem cells exert anti-inflammatory effects on lipopolysaccharide induced human uterine smooth muscle cells effects on lipopolysaccharide induced human uterine smooth muscle cells

Abstract

Mesenchymal stem cells (MSC) possess immunosuppressive properties that mediate their therapeutic potential by integrating into injured tissues and modulating cytokine production. This cell-based therapy has the ability to suppress inflammation in multiple sclerosis, diabetes and graft versus host disease. Inflammation-based models of preterm birth (PTB) use lipopolysaccharide (LPS) to understand mechanisms of disease and investigate potential innovative treatments. TGF-b1 is an anti-inflammatory cytokine can be used as a biomarker to measure response to stimuli. Our objective was to determine whether MSC would suppress an inflammatory response in human uterine cells when exposed to LPS. Human uterine cells were exposed to 5 μg/ml LPS for 4 h. They were then challenged with either 1μg/ml LPS or 100 ng/ml LPS for a subsequent 24 h to elicit an inflammatory response. A minimum concentration of LPS that produced the maximum expression of the pro-inflammatory cytokine IL-6 was determined. Next, co-cultures of uterine cells and MSC were established. After exposing these co-cultures with the optimal dose of LPS, TGF-b1 levels were measured in the supernatant. Human uterine cells and stem cells were obtained from Promocell. Concentrations of IL-6 and TGF-b1 were measured with ELISA. Human uterine cells and MSC were established at a concentration of 200K per well each. The maximum IL-6 expression was 100 pg/ml and occurred at a minimal dose of 100 ng/ml LPS (Fig. 1). When the co-cultures of uterine and MSC were exposed to LPS, there was significantly increased expression of TGF-b1 compared to no LPS, (1215.113 ± 160.0 vs. 807.69 ± 64.0 pg/ml, p<0.0001). IL-6 was not detected in uterine cells without exposure to LPS and TGF-β1 was not detected in the co-culture without LPS. MSC appear to exert an anti-inflammatory response with increased production of TGF-b1 in human uterine cells exposed to LPS. Thus, MSC’S immunomodulatory functions may represent a novel, cell-based therapy for inflammatory etiologies of PTB.View Large Image Figure ViewerDownload Hi-res image Download (PPT)