Investigating The Role of FOXO1 in the Differentiation of Mesenchymal Stem Cells towards Chondrodytes

Abstract

Human mesenchymal stem cells (MSCs) have a profound potential in regenerative medicine. MSCs ability to differentiate into various tissue types in vitro provides a promising approach to tissue engineering and subsequently new clinical treatments. The ultimate and long-term goal of the research of which this thesis is a part of, is to generate a healthy hyaline cartilage that can be implanted in cartilage lesions. In this study the role of FOXO1, a gene significantly up-regulated in early chondrogenic differentiation of MSCs in vitro, was investigated. A three-dimensional scaffold aiming at mimicking the environment in the human body has been constructed for in vitro chondrogenic differentiation of MSCs. FOXO1 has been knocked-down by FOXO1siRNA over the course of 6 days and the effects of the down-regulation on chondrogenic, osteogenic and other relevant genes were analyzed by RT-qPCR and nanostring technology. The results showed that FOXO1 depletion in differentiating MSCs altered the expression of some chondrogenic related genes considerably; however the findings need to be validated further and in multiple donors, as donor variability constitutes an important factor to be considered in result interpretation and conclusion drawing. In addition, FOXO1 has also been over-expressed in MSCs in 2D and the effects of up-regulation on pre-selected genes were analyzed by RT-qPCR. The results show that over-expression of FOXO1 in MSCs has not had an impact on the studied genes.