Project description:We have established that BMP6 is an important endogenous regulator of human osteoblast differentiation. Our preliminary experiment showed that 8 hour BMP6 treatment induced early osteoblast markers in hMSC. In this study, we used microarrays to profile the global gene expression program in hMSC induced by BMP6 treatment and further identify the early osteogenic responses to BMP6 stimulation. Experiment Overall Design: The dataset contains a total of 4 gene chip measurements from duplicate experiments each with paired measurements of human MSC with or without 8 hours BMP6 treatment.
Project description:We have established that BMP6 is an important endogenous regulator of human osteoblast differentiation. Our preliminary experiment showed that 8 hour BMP6 treatment induced early osteoblast markers in hMSC. In this study, we used microarrays to profile the global gene expression program in hMSC induced by BMP6 treatment and further identify the early osteogenic responses to BMP6 stimulation. Keywords: Stress response
Project description:Comparison between extracellular vesicles produced from 2D-cultured human Mesenchymal Stem Cells (hMSC) in starvation, and extracellular vesicles produced from spheroids of hMSC hydrodynamically stimulated in a cross-slot millifluidic chip.
Project description:LncRNA transcriptional profiling of human mesenchymal stem cells comparing control undifferentiated HMSC with Day3 and Day6 adipogenic differentiation stages Three-condition experiment, Day0 vs. Day3 vs Day6 cells. Day0: 4 replicates, Day3: 3 replicates and Day6: 3 replicates.
Project description:LncRNA transcriptional profiling of human mesenchymal stem cells comparing control undifferentiated HMSC with Day3 and Day6 adipogenic differentiation stages
Project description:Background. The regenerative and immunomodulatory properties of human mesenchymal stem cells (hMSC) have raised great hope for their use in cell therapy. However, when intravenously infused, hMSCs fail to reach sites of tissue injury. Fucose addition in α-(1,3)-linkage to terminal sialyllactosamines on CD44 creates the molecule known as hematopoietic cell E-/L-selectin ligand (HCELL), programming hMSC binding to E-selectin that is expressed on microvascular endothelial cells of bone marrow (BM), skin, and at all sites of inflammation. Here we describe how this modification on BM-derived hMSC (BM-hMSCs) can be adapted to good manufacturing practice (GMP) standards. Methods. BM-hMSC were expanded using xenogenic-free media and exofucosylated using α-(1,3)-fucosyltransferases VI (FTVI) or VII (FTVII). Enforced fucosylation converted CD44 into HCELL, and HCELL formation was assessed by western blot, flow cytometry, and cell binding assays. Untreated (unfucosylated), buffer-treated and exofucosylated BM-hMSC were each characterized by immunophenotype , differentiation potential and cell viability and fucosylation stability were assessed at room temperature and at 4°C. Safety was assessed by microbiological testing, karyotype, and c-Myc mRNA expression, and potential effects on genetic reprogramming and in cell signaling were analyzed by gene expression microarrays and receptor tyrosine kinase (RTK) phosphorylation arrays. Results. Our protocol efficiently generates HCELL on clinical-scale batches of BM-hMSC. Exofucosylation yields stable HCELL expression for 48 hours at 4°C. Exofucosylation preserved full cell viability and identity, without changing gene expression or RTK phosphorylation. Discussion. The described exofucosylation protocol using xenogenic-free reagents enforces HCELL expression on hMSC endowing potent E-selectin binding without affecting cell viability or native phenotype. This described protocol is readily scalable for GMP-compliant clinical production.
