In Vitro Mesenchymal Stem Cell Differentiation Landscape
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ABSTRACT:
PROVIDER: PRJNA1006438 | ENA |
REPOSITORIES: ENA
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Project description:We employed a tissue engineered system designed to recapitulate mesenchymal stem cell differentiation and performed single-cell profiling to identify differentiation states. We constructed the Mesenchymal Tissue Landscape (MTL) using single-cell RNA-sequencing (scRNA-seq) data from multiple mesenchymal lineages (osteogenic and adipogenic) at various differentiation stages.
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Project description:During in vitro differentiation, pluripotent stem cells undergo extensive remodeling of their gene expression profiles. While studied extensively at the transcriptome level, much less is known about protein dynamics, which might differ significantly from their mRNA counterparts. Here, we present deep proteome-wide measurements of protein levels during the differentiation of embryonic stem cells.
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Project description:Proteomics was used to explore the effect of metformin on the neural differentiation of gingival mesenchymal stem cells in three-dimensional culture.
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Project description:Differentiation of multipotent mesenchymal stem cells into bone-forming osteoblasts requires strict coordination of transcriptional pathways. Aryl hydrocarbon receptor (AhR) ligands, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), have been shown to alter osteoblast differentiation in vitro and bone formation in multiple developmental in vivo models. The goal of the present study was to establish a global transcriptomic landscape during early, intermediate, and apical stages of osteogenic differentiation in vitro in response to TCDD exposure. Human bone-derived mesenchymal stem cells (hBMSC) were cultured in growth media (GM), osteogenic differentiation media (ODM), or osteogenic differentiation media containing 10 nM TCDD (ODM+TCDD), thus enabling a comparison of the transcriptomic profiles of undifferentiated, differentiated, and differentiated -TCDD-exposed hBMSCs, respectively. In this test system, exposure to TCDD attenuated differentiation of hBMSCs into osteoblasts as evidenced by reduced alkaline phosphatase activity and mineralization. At various timepoints, we observed altered expression of genes that play a role in the Wnt, FGF, BMP/TGF-β developmental pathways, as well as pathways related to extracellular matrix organization and deposition. Reconstruction of gene regulatory networks with the iDREM analysis revealed modulation of transcription factors (TF) including POLR3G, NR4A1, RDBP, GTF2B, POU2F2 and ZEB1, which may putatively influence osteoblast differentiation and the requisite deposition and mineralization of bone extracellular matrix. We demonstrate that the combination of RNA-Seq data in conjunction with the iDREM regulatory model, captures the transcriptional dynamics underlying mesenchymal stem cell differentiation under different conditions in vitro. Model predictions are consistent with existing knowledge and provides a new tool to identify novel pathways and transcription factors that may facilitate a better understanding of the osteoblast differentiation process, perturbation by exogenous agents, and potential intervention strategies targeting those specific pathways.
2023-02-08 | GSE217334 | GEO
Project description:Ability to perform osteogenic differentiation is one of the minimal criteria of mesenchymal stem cells (MSCs). Still, it is generally unknown whether osteogenic differentiation is universal cell fate or various phenotypically similar cell states. Besides this, MSCs and their secretomes are actively using for cell/cell-free therapy development, but systemic inter-source variation in MSCs secretomes, proteomes and differentiation mechanisms are still poorly understood. Therefore, here we compared proteomic and secretomic profiles of human mesenchymal cells from six sources: osteoblasts (bone), WJ-MSCs (Warton’s jelly), AD-MSCs (adipose), PDLSCs (tooth: Periodontal Ligament Stem Cells), DPSCs (tooth: Dental Pulp Stem Cells) and GFs (tooth: Gingival Fibroblasts). For experiments we used cells in early passages (3-5) isolated from 3-6 individuals. All cells were compared in standard cultivation and in the 10th day after induction of osteogenic differentiation.
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Project description:Adult neural stem cells (aNSCs) show multilineage differentiation potential influenced by intrinsic and extrinsic signaling cues. We and others have shown that stimulation of aNSCs with bone marrow mesenchymal stem cell (MSC) secreted factors substantially enhances in vitro oligodendrogenesis at an expense of astrogenesis by yet unknown mechanisms (Rivera et al. 2006, Jadasz et al. 2013; 2018, Rivera et al., 2019). In the present study, we demonstrate that aNSCs pre-treated with MSC secretomes for different periods in vitro preferentially differentiate to oligodendrocytic cells in vivo after transplantation into the adult rat spinal cord. Analysis of different time points after transplantation revealed a stable survival rate of transplanted aNSCs and an emphasized pro-oligodendroglial differentiation in response to MSC secreted factors. MSC derived secretomes were then analyzed by mass spectrometry-based proteomics and label-free quantification to identify secreted proteins contributing to oligodendroglial lineage fate determination. To exclude possible contaminants derived from dead cells or serum, our approach includes a comparison of the abundances of proteins present in MSC derived secretomes with corresponding proteins in cell lysates (Grube et al., 2018, Schira-Heinen et al., 2019).
2020-06-18 | PXD018231 | Pride
Project description:One of the promising approaches to slow down or treat neurodegenerative diseases or spinal cord injuries represent cell therapies. The grafted cells could either integrate into the damaged tissue to replace dead or damaged cells or by secretion of different factors modulate inflammatory reaction, reduce tissue damage and support neuronal survival. However, due to the heterogeneity of in vitro cultured cells, comprehensive characterization of such cells is absolutely crucial to prevent safety risks.
Here, we performed SWATH-MS analysis to characterize changes in proteome of human neural stem cells (NSCs) during their differentiation either spontaneously by withdrawal of EGF and FGF2 in cell culture media or by trophic support of BDNF/GDNF growth factors. We quantified about 2800 proteins over the 28 days of differentiation and showed that changes in cellular proteomes are caused mostly by differentiation time course, rather than type of differentiation itself and that the major changes in protein expression occurred between day 0 and day 7 of both differentiations.
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