Project description:The columnar epithelial cells comprising the intestinal tract, stomach, and uterus can be cultured in vitro as organoids or in adherent culture. However, the proliferation of these columnar epithelial cells in adherent culture is limited. Likewise, human pluripotent stem cell (hPSC)-derived intestinal epithelial cells do not show extensive or clonal propagation in vitro. In this study, we induced proliferation of hPSC-derived small intestinal epithelium for a longer time by utilizing mesenchymal stromal cells derived from self-organized intestinal organoids as feeders. The proliferating cells exhibited columnar form, microvilli and glycocalyx formation, and cell polarity, as well as expression of drug-metabolizing enzymes and transporters. It is noteworthy that small intestinal epithelial stem cells cannot be cultured in adherent culture alone, and the stromal cells cannot be replaced by other feeders. Organoid-derived mesenchymal stromal cells resemble the trophocytes essential for maintaining small intestinal epithelial stem cells and play a crucial role in adherent culture. The high proliferative expansion, productivity, and functionality of hPSC-derived small intestinal epithelial stem cells could have potential applications in pharmacokinetic and toxicity studies and regenerative medicine.
Project description:Stromal-epithelial interactions play a fundamental role in tissue homeostasis, controlling cell proliferation and differentiation. Not surprisingly, aberrant stromal-epithelial interactions contribute to malignancies. The goals and objectives of this study were 1.) to characterize and validate the molecular identity of human primary epithelial and stromal/mesenchymal breast cells maintained long-term in novel ex vivo culture conditions in serum free medium. 2.) To analyze changes in gene expression profiles of normal human primary epithelial and stromal/mesenchymal breast cells upon long-term ex vivo co-culture when compared to corresponding monocultures 3.) To study the dynamic reciprocity between normal human primary epithelial and stromal/mesenchymal breast cells. 4.) To identify critical molecular pathways and biomarkers controlling epithelial and/or stromal cell growth and quiescence. Human primary epithelial progenitor cells and mesenchymal stem cells bearing fluorescent tags were either co-cultured in novel ex vivo culture conditions on ECM coated meshes in serum free medium (M5) or cultured as monocultures in the same conditions for 30 days. The cultures were then dissociated and epithelial and stromal/mesenchymal cells from either co-cultures or monocultures separated by FACS. Gene expression profiling of epithelial or stromal/mesenchymal cells was performed. Clean gene expression profiles from three different epithelial and stromal/mesenchymal cell extracts either grown in co-cultures or monocultures were successfully obtained.
Project description:Growing evidence indicates that tumor-associated stroma plays a negative role in human colorectal cancer (CRC). Nature of specific stromal cell populations involved and mechanisms underlying their negative impact remain to be fully understood. In this study we describe the expansion from human primary CRCs of a mesenchymal cell population, referred to as tumor-associated stromal cells (TASCs), resembling bone marrow-derived mesenchymal stem cells (BM-MSCs) in morphology, phenotypes and differentiation potential. We found that, upon co-culture with tumor cells, TASCs acquire membrane-bound TGF-mbTGF-expression, a phenomenon mediated by v6 integrin. MbTGF-expression proved to be critical for triggering epithelial-to-mesenchymal transition (EMT) in tumor cells, eventually leading to enhanced dissemination of circulating tumor cells and increased metastasis formation, in an orthotopic mouse model. Our data identify CRC-associated mesenchymal stem-like cells as critical EMT initiators and suggest mbTGF- as potential novel therapeutic target.
Project description:Metabolomics and lipidomics workflows were used to analyze Mesenchymal stromal cell (MSC) metabolites. Metabolite abundances were used to model MSC potency results in IDO and T-cell proliferation assays.
Project description:The raw files of the proteomics dataset are N=18 and a brief description of the files is shown below:
AF1 raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 1
AF2.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 2
AFA1.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 3
AFA2.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 4
AFB1.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 5
AFB2.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 6
HL1A.raw: exosomes derived from Hepatocyte like cells 1
HL1B.raw: exosomes derived from Hepatocyte like cells 2
HL2A.raw: exosomes derived from Hepatocyte like cells 3
HL2B.raw: exosomes derived from Hepatocyte like cells 4
HL3A.raw: exosomes derived from Hepatocyte like cells 5
HL3B.raw: exosomes derived from Hepatocyte like cells 6
HPLA1.raw: exosomes derived from Hepatic Progenitor-like cells 1
HPLA2.raw: exosomes derived from Hepatic Progenitor-like cells 2
HPLB1.raw: exosomes derived from Hepatic Progenitor-like cells 3
HPLB2.raw: exosomes derived from Hepatic Progenitor-like cells 4
HPLA.raw: exosomes derived from Hepatic Progenitor-like cells 5
HPLB.raw: exosomes derived from Hepatic Progenitor-like cells 6
Raw files analysis was performed with Proteome Discoverer 1.4 (Thermo) software package, using the Sequest search engine and the Uniprot human (Homo sapiens) reviewed database, downloaded on December 15, 2017, including 20,243 entries. The search was performed using carbamidomethylation of cysteine as static and oxidation of methionine as dynamic modifications. Two missed cleavage sites, a precursor mass tolerance of 10 ppm and fragment mass tolerance of 0.05 Da were allowed. False discovery rate (FDR) validation was based on q value: target FDR : 0.05. Label free quantification was performed by utilizing the precursor ion area values exported from the total ion chromatogram as defined by the Proteome Discoverer v. 1.4.0.288 (Thermo Scientific). Output files from Proteome Discoverer were processed with R programming language for statistical computing (version 4.0.3).
The following comparisons were made: AF vs HL, AF vs HPL and HL vs HPL.
The msf files are N=18 and have the same name as the raw files above.
Project description:<p><strong>BACKGROUND:</strong> Ischemia/reperfusion injury (IRI) is the leading cause of acute kidney injury (AKI). The current standard of care focuses on supporting kidney function, stating the need for more efficient and targeted therapies to enhance repair. Mesenchymal Stromal Cells (MSCs) and their secretome, either as conditioned medium (CM) or extracellular vesicles (EVs), have emerged as promising options for regenerative therapy, however, their full potential in treating AKI remains unknown.</p><p><strong>METHODS:</strong> In this study, we employed an in vitro model of chemically-induced ischemia using antimycin A combined with 2-deoxy-D-glucose to induce ischemic injury in proximal tubule epithelial cells. Afterwards, we evaluated the effects of MSC secretome, CM or EVs obtained from adipose tissue, bone marrow and umbilical cord, on ameliorating the detrimental effects of ischemia. To assess the damage and treatment outcomes, we analyzed cell morphology, mitochondrial health parameters (mitochondrial activity, ATP production, mass and membrane potential) and overall cell metabolism by metabolomics.</p><p><strong>RESULTS:</strong> Our findings show that ischemic injury caused cytoskeletal changes confirmed by disruption of the F-actin network, energetic imbalance as revealed by a 50% decrease in the oxygen consumption rate, increased oxidative stress, mitochondrial dysfunction and reduced cell metabolism. Upon treatment with MSC secretome, the morphological derangements were partly restored and ATP production increased by 40-50%, with umbilical cord-derived EVs being most effective. Furthermore, MSC treatment led to phenotype restoration as indicated by an increase in cell bioenergetics, including increased levels of glycolysis intermediates, as well as an accumulation of antioxidant metabolites.</p><p><strong>CONCLUSION:</strong> Our in vitro model effectively replicated the in vivo-like morphological and molecular changes observed during ischemic injury. Additionally, treatment with MSC secretome ameliorated proximal tubule damage, highlighting its potential as a viable therapeutic option for targeting AKI.</p>
Project description:This SuperSeries is composed of the following subset Series: GSE30064: Cultured human amniotic fluid-derived mesenchymal stromal cells [PIQOR data] GSE30065: Cultured human amniotic fluid-derived mesenchymal stromal cells [miRXplore data] Refer to individual Series
Project description:Mesenchymal stromal cells (MSCs) have been shown to exert their therapeutic effects through the secretion of various paracrine factors, including extracellular vesicles (EVs). These EVs are now being developed as a promising alternative to cell-based therapies. Menstrual blood-derived stromal cells (MenSCs) are a type of MSC that have emerged as a innovative source due to their immunomodulatory and regenerative properties. Additionally, new strategies of cell priming could potentially alter the concentration and cargo of released EVs, leading to modifications in their biological properties. In this study, we aimed to characterize the EVs released by MenSCs and to compare their therapeutic potential under three different preconditioning conditions (proinflammatory stimuli, physioxia, and acute hypoxia).