Project description:To investigate the function TWIST1 in the regulation of stemness in mesenchymal stem cell, we established human mesenchymal stem cell in TWIST1-overexpression.
Project description:To investigate the function TWIST1 in the regulation of stemness in mesenchymal stem cell, we established human mesenchymal stem cell in TWIST1-overexpression.
Project description:To investigate the effects of TWIST1 expression on mesenchymal stem cells, we established a retrovirally transduced MSC line overexpressing TWIST1.
Project description:Mesenchymal stem/stromal cells (MSCs) function as skeletal progenitors in bone marrow and regulate hematopoiesis and hemodynamic processes via secretion of paracrine acting factors. Although TWIST1 plays important roles in mesoderm formation and skull and vascular development, its role in MSCs is poorly defined. Therefore, we conducted ChIP-seq analysis to identify genes potentially regulated by TWIST1.
Project description:It is well known that the properties of hematopoietic stem cells (HSCs) such as their ability to self-renew, and multipotency are maintained through interactions with mesenchymal cells. Meanwhile, the identification of cells that are the key component of niche remains a subject of dispute. Mesenchymal stem cells (MSCs) are one of the niche component cells, MSCs are one of the niche component cells which provide stemness signal to HSCs, while it is also hypothesized that there is also a counterpart signal from hematopoiesistic cells to mesenchymeal cells, as HSCs also constitute core stem cells niche for MSCs. Although mesenchymal stem cells (MSCs) have also been studied intensively, the signals involved in the maintenance of their properties have not attracted much attentions. We used microarrays to detail the global programme of gene expression underlying HSC-MSC interaction and identified distinct classes of regulated genes during this process.
Project description:In addition to their stem/progenitor properties, mesenchymal stem cells (MSCs) also exhibit various effector functions potent effector (angiogenic, anti-inflammatory, immune-modulatory) functions that are largely paracrine in nature. It is widely believed that effector functions underlie most of the therapeutic potential of MSCs and are independent of their stem/progenitor properties. Here we demonstrate that stem/progenitor and effector functions are coordinately regulated at the cellular level by the transcription factor Twist1 and specified within populations according to a hierarchical model. We further show that manipulation of Twist1 levels by genetic approaches or by exposure to widely used culture supplements including fibroblast growth factor 2 (Ffg2) and interferon gamma (IFN-gamma) alters MSC efficacy in cell-based and in vivo assays in a predictable manner. Thus, by mechanistically linking stem/progenitor and effector functions our studies provide a unifying framework in the form of an MSC hierarchy that models the functional complexity of populations. Using this framework, we developed a Clinical Indications Prediction (CLIP) scale that predicts how donor-to-donor heterogeneity and culture conditions impact the therapeutic efficacy of MSC populations for different disease indications. We used microarrays to detail the global gene expression in response to genetic and growth factor manipulation of TWIST1 expression and function. Mesenchymal stem cells at subconfluent culture in growth media (CONTROL), or supplemented with FGF2 (20 ng/ml) (FGF2), trasfected with scrambled (SCRAMBLED) or TWIST1 siRNA (TWIST1) were used for RNA extraction and hybridization on Affimentrix microarrays. We pooled three independently extracted RNA samples per group.