Acute respiratory distress syndrome (ARDS) is an acute and life-threatening inflammatory condition that affects the lungs and is predominantly caused by alveolar injury and/or damage. This results in the release of pro-inflammatory cytokines and subsequent neutrophil recruitment to the lungs. Neutrophil activation in response to pro-inflammatory stimuli causes the release of cytotoxic compounds that further damages the surrounding capillary and alveolar epithelium. Mesenchymal stromal cells (MSC) are bone marrow-derived cells that demonstrate pro-reparative and immunomodulatory effects in vitro and in vivo. The hypothese of this research project aims to investigate the influence of ARDS patient serum (from blood) on the cytoprotective ability of MSCs in vitro.The aims of this project are to examine the ability of MSCs to promote wound healing in epithelial cells in vitro in the presence of serum from ARDS patients and to investigate the influence of ARDS serum on MSCs function in vitro.
The first part of this project’s methodology will be the preparation of aseptic culture of MSCs and the aseptic culture of airway epithelial cells in vitro. MSCs will be exposed to hypoinflammatory or hyperinflammatory ARDS serum to simulate exposure to the ARDS patient microenvironment. Conditioned medium (CM) will be collected from the ARDS patient environment exposed MSCs. After exposure to the ARDS patient microenvironment, the capacity for MSC CM to enhance wound healing will be investigated. Additionally, MCSs ability to promote wound healing will be performed using a wound healing assay in epithelial cells.
The second part of this project project’s methodology will be to investigate the influence of ARDS serum on MSC activation and production of cytoprotective factors (VEGF, HGF, ANGPTL4). Expression of these proteins will be examined by ELISA. Thorough and accurate recording and reporting of experimental practices will all be included in the final powerpoint and word document.