Project description:The classical concept of bone marrow-derived mesenchymal stem cells (BM-MSC), intended as a uniform, broad potent population, is progressively being substituted by the idea that the bone marrow harbors heterogeneous populations of non-hematopoietic stem cells. This in vivo heterogeneity is also amplified by the different experimental strategies used to isolate/culture them. Among the exogenous factors described to affect MSC in vitro growth, basic-fibroblast growth factor (bFGF) is one of the most common growth factors used to expand stem cells. Moreover, it has been reported that its signaling is associated with the mainteinance of stemness of a variety of stem cells, included MSC. Using an ectopic model of bone regeneration, we have previously described that the implantation of cells with different commitment levels, differentially influences the capacity to recruit host cells, activating endogenous regenerative mechanisms. Due to its properties, we here demonstrate that the addition of bFGF to primary BM cultures, leads to the selection of specific subpopulations able to induce a different host regenerative response, when in vivo implanted in association with suitable ceramic scaffolds. Moreover, taking advantage of a multiparametric and comparative genomic and proteomic approach, it has been evaluated how different culture conditions combine to bring about appreciable changes in the secretome of the cells, that consequently influence their in vivo regenerative behaviour. The full comprehension of the regulatory mechanisms that rule the host response depending on the type and differentiative stage of the transplanted cells could help us to develop novel clinical strategies where host cells could directly contribute to regenerate the appropriate tissue. Comparison of bFGF effects on bone marrow mesenchymal stem cells.
Project description:The classical concept of bone marrow-derived mesenchymal stem cells (BM-MSC), intended as a uniform, broad potent population, is progressively being substituted by the idea that the bone marrow harbors heterogeneous populations of non-hematopoietic stem cells. This in vivo heterogeneity is also amplified by the different experimental strategies used to isolate/culture them. Among the exogenous factors described to affect MSC in vitro growth, basic-fibroblast growth factor (bFGF) is one of the most common growth factors used to expand stem cells. Moreover, it has been reported that its signaling is associated with the mainteinance of stemness of a variety of stem cells, included MSC. Using an ectopic model of bone regeneration, we have previously described that the implantation of cells with different commitment levels, differentially influences the capacity to recruit host cells, activating endogenous regenerative mechanisms. Due to its properties, we here demonstrate that the addition of bFGF to primary BM cultures, leads to the selection of specific subpopulations able to induce a different host regenerative response, when in vivo implanted in association with suitable ceramic scaffolds. Moreover, taking advantage of a multiparametric and comparative genomic and proteomic approach, it has been evaluated how different culture conditions combine to bring about appreciable changes in the secretome of the cells, that consequently influence their in vivo regenerative behaviour. The full comprehension of the regulatory mechanisms that rule the host response depending on the type and differentiative stage of the transplanted cells could help us to develop novel clinical strategies where host cells could directly contribute to regenerate the appropriate tissue.
Project description:Data files associated with the manuscript titled "Development of MDM2-targeting PROTAC for Advancing Bone Regeneration," which is currently under review. This manuscript introduces MDM2-targeting PROTACs customized for application in bone regeneration. We developed MDM2-PROTACs (CL144) that demonstrated potent degradation efficiency and a strong inductive effect on biomineralization. Proteomics analysis was performed on human bone marrow-derived mesenchymal stem cells (hBMSCs) to investigate the degradation selectivity of the compound. Through proteomics database searches, we identified 6,388 proteins, including several differentially expressed proteins (DEPs), many of which are known to interact with MDM2 or play significant roles in the ubiquitin-proteasome system.
Project description:Normal SJL mice, 6 to 8 weeks old, were used for the isolation of bone marrow stem cells (BMSC). Bone marrow cells were obtained from the femurs and tibias of euthanized mice by flushing with PBS. Cells were subjected to negative magnetic sorting using the Lineage Cell Depletion Kit. Isolation of murine bone marrow Lin-Sca-1+ cells was performed using MACS Sca-1 MultiSort Kit (fBMSC). The differentiation of neural stem cells was induced by removing the bFGF-containing medium and resuspending cells in fresh bFGF-free medium. Microarray analysis of miRNA expression profiles was performed comparing non differentiated bone marrow stem cells (fBMSC) to bone marrow stem cells differentiated for 4 or 7 days.
Project description:Mesenchymal stem cells (MSCs)-derived exosomes (exo) have shown comprehensive application prospects over the years. Despite similar functions, exomes from different origins present heterogeneous characteristics and components; however, there are no relevant proteomic analyses. In this study, we isolated exosomes from MSCs, derived from different tissues, by ultracentrifugation. A total of 1014 proteins were detected using a label-free method and analyzed with bioinformatics tools. The results revealed their shared function in the extracellular matrix receptor. Bone marrow-MSCs-derived exosomes showed superior regeneration ability. Likewise, adipose tissue-MSCs-derived exosomes played a significant role in immune regulation. Whereas, umbilical cord-MSCs-derived exosomes were more prominent in tissue damage repair.
Project description:2D IDA protein quantitation of mesenchymal stem cells derived from bone
marrow across five donors. A total of 10 2D LC-MS runs were performed, using cells both not stimulated and following a 20 hour treatment with interferon gamma.