Project description:Global DNA methylation profiling of hepatocyte-like cells. We established a protein-based artificial transcription system and successfully engineered human hepatocyte-like cells from human umbilical cord-derived mesenchymal stem cells (MSC-UCs). We utilized four artificial transcription factors (ATFs) that targeted hepatocyte nuclear factor (HNF)1α, HNF3γ, HNF4α and GATA4 (4F), and found that treatment with 4F for 3 (4Fd3) or 5 (4Fd5) days converted MSC-UCs to hepatocyte-like cells. We then examined global DNA methylation profiles of 4Fd3, 4Fd5, NTP-EGFP protein treated cells (NE) and non-treated cells (NC).
Project description:We established a protein-based artificial transcription system and successfully engineered human hepatocyte-like cells from human umbilical cord-derived mesenchymal stem cells (MSC-UCs). We utilized four artificial transcription factors (ATFs) that targeted hepatocyte nuclear factor (HNF)1α, HNF3γ, HNF4α and GATA4 (4F), and found that treatment with 4F for 5 days converted MSC-UCs to hepatocyte-like cells (4F-Heps). We then examined gene expression profiles of 4F-Heps and compared them with those of non-treated MSC-UCs (NC) or MSC-UCs treated with NTP-EGFP protein as control protein (NE).
Project description:Comparison of whole genome gene expression profiles of stem cells from exfoliated decidous teeth (SHEDs) ,Hepatocyte like-cells derived SHEDs (SHED-Hep cells), and human primary hepatocytes. We showed that the profile of three different donors of MSC-Hep cells was close to that of human hepatocytes, but separate from that of three different donors of undifferentiated MSCs
Project description:Mesenchymal stromal cells (MSC) are ideal candidates for cell therapies, due to their immune-regulatory and regenerative properties. We have previously reported that lung-derived MSC are tissue-resident cells with lung-specific properties compared to bone marrow-derived MSC. Assessing relevant molecular differences between lung-MSC and bone marrow-MSC is important, given that such differences may impact their behavior and potential therapeutic use. Here, we present an in-depth mass spectrometry (MS) based strategy to investigate the proteomes of lung-MSC and bone marrow-MSC. The MS-strategy relies on label free quantitative data-independent acquisition (DIA) analysis and targeted data analysis using a MSC specific spectral library. We identified several significantly differentially expressed proteins between lung-MSC and bone marrow-MSC within the cell layer (352 proteins) and in the conditioned medium (49 proteins). Bioinformatics analysis revealed differences in regulation of cell proliferation, which was functionally confirmed by decreasing proliferation rate through Cytochrome P450 stimulation. Our study reveals important tissue-specific differences within proteome and matrisome profiles between lung- and bone marrow-derived MSC that may influence their behavior and affect the clinical outcome when used for cell-therapy.
Project description:Mesenchymal stem cells (MSC) have emerged as potent therapeutic tool for a number of pathologies, including immune ones. However, unwelcome effects of MSC on the blood coagulation were revealed in some cases, which require more in-depth analysis. In this study, we explored the trombotic properties of human MSC from umbilical cord. We revealed strong procoagulant effects of umbilical cord MSC toward human and rat whole blood and platelets-free plasma using rotational thromboelastometry and thrombodynamics tests. The similar potentiation of clotting was demonstrated for MSC-derived extracellular vesicles (EV). In order to suggest approaches to avoid unwanted effects we studied the impact of heparin supplement on MSC/EV procoagulation properties. We found that therapeutic doses of unfractionated heparin injected in the patient's blood (administered in vivo) did not abrogate the procoagulant properties of MSC. Mass-spectrometry analysis of proteins of MSC and EV involved in coagulation-associated pathways was used to evaluate mechanisms of protrombotic effects.
Project description:Genome-wide analysis of human bone marrow-derived MSC cells in comparision with MSC with EGR1 knockdown or EGR1 overexpressing cells