Project description:Quantitative shotgun proteomic analysis (TMT) of the effect of inhibition of MIR21 in the EV protein cargo of human, Umbilical cord-derived Mesenchymal Stem Cells.
Project description:This study investigated early host reactions to implanted materials to predict successful tissue regeneration with implant. Three kinds of scaffold, i.e., non-coat, collagen-coated, and PMB-coated porous polystylene scaffolds were implanted subcutaneously in mice dorsal area. Those scaffolds were used as bio-incomopatible materials, appropriate materials for tissue regeneration (bio active), and inappropriate to regenration (bio-inert) scaffolds. Seven days after implantation, scaffolds were explanted and total RNA was isolated from infiltrated host cells into scaffold by laser microdissection. Gene expressions of cells in collagen- and PMB-coated scaffold were normalized using results of non coat scaffold. Genes with more than 2-fold difference between collagen and PMB were picked up and narrowed to related keywords; inflammation, angiogenesis, wound healing, and mcrophage polarization. Among those genes, interluekin (IL)-1beta which promote both inflammation and wound healing was up-regulated in collagen-coated scaffold. On the other hand, IL-10 which suppress both inflammation and wound healing was up-regulated in PMB-coated scaffold. Angiogenesis-promoting genes were up-regulated and angiogenesis suppressve genes were suppressed in collagen. Up-regulation of IL-1b and the angiogenesis-relating genes inside the porous scaffolds are the possibly important factors for controlling tissue regeneration. Three-condition experiment, host cells infiltrated in non coat (reference), collagen-coated, and PMB-coated scaffolds. Two-microarray condition experiments, collagen vs. non coat and PMB coat vs. non coat. Hybridization: 2 replicates. Scanning: 3 replicates. Biological experiments: once.
Project description:Human umbilical cord mesenchymal stem cells maintained multipotency and immunosuppressive ability when being cultured in chemical defined serum free medium, but gained different gene expression profile. We used microarrays to identify the transcriptional difference between human umbilical cord mesenchymal stem cells cultured in serum containing medium and chemical defined serum free medium. human umbilical cord mesenchymal stem cells were cultured in conventional serum containing medium and chemical defined serum free medium separately. Total RNA was extracted and hybridized on Affymetrix microarrays.
Project description:A genomic expression comparison was done among neural progenitor cells cultured on 2D substrates, 3D porous polystyrene scaffolds, and as 3D neural spheres (in vivo surrogate), with the goal of assessing the feasibility of establishing the meaning of 3D and associated physiological relevance at the molecular level Neural progenitor cells were cultured on 2D surfaces, in 3D scaffolds and as 3D neural spheres. Chemical cues are controlled by coating. Only spacial properties of the culture systems were compared.
Project description:A transcriptomic expression comparison was done among superior cervical ganglion (SCG) cells cultured on 2D substrates, 3D porous polystyrene scaffolds, and in freshly dissected tissue (in vivo surrogate), with the goal of assessing the feasibility of establishing the meaning of 3D and associated physiological relevance at the molecular level SCG cells were cultured on 2D surfaces and in 3D scaffolds. Chemical cues are controlled by coating. Only spacial properties of the culture systems were compared. Cells from freshly dissected tissue were used as in vivo surrogates for positive control of 3D cells.
Project description:This study investigated early host reactions to implanted materials to predict successful tissue regeneration with implant. Three kinds of scaffold, i.e., non-coat, collagen-coated, and PMB-coated porous polystylene scaffolds were implanted subcutaneously in mice dorsal area. Those scaffolds were used as bio-incomopatible materials, appropriate materials for tissue regeneration (bio active), and inappropriate to regenration (bio-inert) scaffolds. Seven days after implantation, scaffolds were explanted and total RNA was isolated from infiltrated host cells into scaffold by laser microdissection. Gene expressions of cells in collagen- and PMB-coated scaffold were normalized using results of non coat scaffold. Genes with more than 2-fold difference between collagen and PMB were picked up and narrowed to related keywords; inflammation, angiogenesis, wound healing, and mcrophage polarization. Among those genes, interluekin (IL)-1beta which promote both inflammation and wound healing was up-regulated in collagen-coated scaffold. On the other hand, IL-10 which suppress both inflammation and wound healing was up-regulated in PMB-coated scaffold. Angiogenesis-promoting genes were up-regulated and angiogenesis suppressve genes were suppressed in collagen. Up-regulation of IL-1b and the angiogenesis-relating genes inside the porous scaffolds are the possibly important factors for controlling tissue regeneration.
Project description:As an important part of regenerative medicine, human umbilical cord mesenchymal stem cells show a good clinical therapeutic effect, but their use is still limited. In recent years, the use of umbilical cord mesenchymal stem cell exosomes as cell replacement therapy can effectively overcome some defects of cell therapy. However, whether there are differences among different batches of exosomes, the specific mechanism of exosomes intervention is still poorly known. In this study, LC-MS/MS was used to identify the protein composition of two generations exosomes from three different donors, and the function and possible mechanism of exosome proteomic of human umbilical cord mesenchymal stem cells was analyzed by bioinformatics. It was found that the protein composition of human umbilical cord mesenchymal stem cell exosomes was basically the same in 6 groups, and 676 core proteins were found. The biological function of core proteomic was analyzed by GO, and it was found that core proteomic was involved in 88 molecular functions, such as anion binding, nucleotide binding, receptor binding, ribonucleotide binding; 648 biological processes, such as regulation of cellular process, macromolecule metabolic process, transport; 157 cellular components. The regulation pathway of core proteomic was analyzed by KEGG, and it was found that the regulation of blood coagulation, bacterial infection, phagocytosis, vesicle circulation and so on. Umbilical cord mesenchymal stem cell exosomes were used to interfere with APP/PS1 transgenic mice to explore the mechanism of exosome regulation of synaptic vesicle cycle signal pathway in Alzheimer's disease. The results showed that the exosomes could significantly enhance the spatial memory and learning ability, exercise ability and anti-fatigue ability of Alzheimer's disease model mice. Further analysis of mouse hippocampal proteome showed that the exosome proteomic of human umbilical cord mesenchymal stem cells was enriched into 9 proteins of synaptic vesicle cycle signal pathway. Compared with control group and exosome group, the contents of AP2A1 and AP2B1 in hippocampus of model group were significantly decreased. The results of this study can provide research methods and theoretical basis for the use of human umbilical cord mesenchymal stem cell exosomes to treat diseases, and further promote its clinical application.