Mesenchymal stem cells (MSCs) hold therapeutic potential for a number of pathologic conditions due to their ability to differentiate into various cell types. However, the clinical application of MSCs requires them to be maintained in a non-differentiated state, which remains a major challenge for the field. To develop ready-to-go MSCs, specific culture conditions that maintain the ‘stemness’ of the MSCs need to be formulated. In this study, the ‘stemness’ of rat MSCs cultured under six separate culture conditions is being investigated using a proteomics based approach.
Bone marrow derived rat MSCs were established in standard culture medium. Cells were then serum starved prior to culture for 72 hours in standard growth media supplemented with 2 % serum and a combination of fibronectin (FN), fibroblast growth factor 2 (FGF2) and/or bone morphogenetic protein 4 (BMP4). Cellular protein was collected and prepared using standard techniques for qualitative and quantitative (SWATH) mass spectrometry.
From this study, a library of all detectable proteins in rat MSCs was generated. Each protein was quantified across the culture conditions and the predominant biological processes were determined. Fibronectin was determined to have a significant role on the biology of the cells. Furthermore, statistical analysis of the quantitative profiles of each of the treatment groups revealed eight specific proteins that were of interest across the culture condition.
Characterisation of the rat MSC proteome provides insight into the molecular processes that occur within these cells while in a ‘stem’ state, allowing for the development of defined culture conditions that maintain MSCs in an undifferentiated state.
This study will provide much needed information on culture conditions that enable monitoring and maintenance of MSCs in a non-differentiated state, which in the future could permit an off-the-shelf MSC therapeutic to be developed for the treatment of a variety of medical conditions.