Development of a human adipocyte model derived from human mesenchymal stem cells (hMSC) as a tool for toxicological studies on the action of TCDD

Abstract Efforts were made to develop a human adipocyte model that is useful for toxicological studies in vitro. For this purpose, a stem cell line derived from human bone marrow cells, originally from an adult, was induced to differentiate towards adipocytes by treating them with insulin, dexamethasone, indomethacin and 3-isobutyl-1-methylxanthine for 3 d, followed by additional incubation for 3 d in Dulbecco's modified Eagle's medium supplemented with insulin only. In most cases, thus differentiated cells through such one cycle of differentiation treatment were further subjected to the second cycle of differentiation. The resulting 2-cycle differentiated cells were found to exhibit many characteristics of typical adipocytes. Dioxin (TCDD), when added at the beginning of their treatment with differentiation-inducing hormone cocktail, clearly prevented them from becoming adipocytes, as in the case of TCDD-treated 3T3-L1 cells. Furthermore, TCDD, even when administered to previously differentiated human mesenchymal stem cells (hMSC) adipocytes, consistently induced the sign of inflammatory responses during the early period of TCDD action (24 h), which was followed by gradual loss of adipocyte-specific markers during the 5-d incubation period. In conclusion, hMSC-derived adipocytes appear to offer a promising human cell model suited for future toxicological studies.

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Induced neural differentiation of human mesenchymal stem cells affects lipid metabolism pathways

10.1101/2021.01.17.427010 ◽

2021 ◽

Author(s):

Pnina Green ◽

Inna Kan ◽

Ronit Mesilati-Stahy ◽

Nurit Argov-Argaman ◽

Daniel Offen

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Neuronal Development ◽

Human Mesenchymal Stem Cells ◽

Fold Increase ◽

Fatty Acid Binding Proteins ◽

Prostaglandin E ◽

Fatty Acid Binding ◽

Differentiated Cells ◽

Neuronal Membranes

AbstractNeuronal membranes contain exceptionally high concentrations of long-chain polyunsaturated fatty acids (PUFA), docosahexaenoic acid (DHA) and arachidonic acid (ARA), which are essential for neuronal development and function. Adult bone-marrow-derived mesenchymal stem cells (MSC) can be induced to possess some neuronal characteristics. Here we examined the effects of neuronal induction on the PUFA metabolism specific pathways. Differentiated cells contained ~30% less ARA than MSC. The expression of specific ARA metabolizing enzymes was upregulated, notably that of prostaglandin E2 synthase which increased more than 15-fold, concomitantly with a 3-fold increase in the concentration of PGE2 in the medium. Moreover, induced differentiation was associated with enhanced incorporation of exogenous DHA, upregulation of acyl-CoA synthases, fatty acid binding proteins, choline kinase (CK) and phosphatidylserine synthases as well as increased total cellular phospholipids (PL). These findings suggest that active ARA metabolites may be important in the differentiation process and that neuronal induction prepares the resulting cells for increased DHA incorporation through the action of specific enzymes.

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Regulation of CXCR6 Expression on Adipocytes and Osteoblasts Differentiated from Human Adipose Tissue-Derived Mesenchymal Stem Cells

Stem Cells International ◽

10.1155/2020/8870133 ◽

2020 ◽

Vol 2020 ◽

pp. 1-14

Author(s):

Seung-Cheol Lee ◽

Yoo-Jung Lee ◽

Min Kyoung Shin ◽

Jung-Suk Sung

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Cell Activation ◽

Molecular Mechanisms ◽

De Novo ◽

Human Mesenchymal Stem Cells ◽

De Novo Synthesis ◽

Differentiation Potential ◽

Differentiated Cells

Human mesenchymal stem cells derived from adipose tissue (hADMSCs) are a desirable candidate in regenerative medicine. hADMSCs secrete growth factors, cytokines, and chemokines and also express various receptors that are important in cell activation, differentiation, and migration to injured tissue. We showed that the expression level of chemokine receptor CXCR6 was significantly increased by ~2.5-fold in adipogenic-differentiated cells (Ad), but not in osteogenic-differentiated cells (Os) when compared with hADMSCs. However, regulation of CXCR6 expression on hADMSCs by using lentiviral particles did not affect the differentiation potential of hADMSCs. Increased expression of CXCR6 on Ad was mediated by both receptor recycling, which was in turn regulated by secretion of CXCL16, and de novo synthesis. The level of soluble CXCL16 was highly increased in both Ad and Os in particular, which inversely correlates with the expression on a transmembrane-bound form of CXCL16 that is cleaved by disintegrin and metalloproteinase. We concluded that the expression of CXCR6 is regulated by receptor degradation or recycling when it is internalized by interaction with CXCL16 and by de novo synthesis of CXCR6. Overall, our study may provide an insight into the molecular mechanisms of the CXCR6 reciprocally expressed on differentiated cells from hADMSCs.

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MP098FACTORS RELEASED BY HUMAN MESENCHYMAL STEM CELLS SUPPRESS GLUCOSE-INDUCED INFLAMMATORY RESPONSES OF STABLE RENAL PROXIMAL TUBULAR EPITHELIAL CELL MONOLAYERS

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfx162.mp098 ◽

2017 ◽

Vol 32 (suppl_3) ◽

pp. iii461-iii462

Author(s):

Md Nahidul Islam ◽

Tomás Griffin ◽

Elizabeth Sander ◽

Joana Cabral ◽

Thomas Ritter ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Epithelial Cell ◽

Inflammatory Responses ◽

Human Mesenchymal Stem Cells ◽

Tubular Epithelial Cell ◽

Cell Monolayers ◽

Proximal Tubular ◽

Epithelial Cell Monolayers ◽

Renal Proximal Tubular

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Menaquinone-7 Supplementation Improves Osteogenesis in Pluripotent Stem Cell Derived Mesenchymal Stem Cells

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2020.618760 ◽

2021 ◽

Vol 8 ◽

Author(s):

Asim Cengiz Akbulut ◽

Grzegorz B. Wasilewski ◽

Nikolas Rapp ◽

Francesco Forin ◽

Heike Singer ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Human Mesenchymal Stem Cells ◽

Cellular Migration ◽

Collagen Deposition ◽

Runx2 Expression ◽

Differentiated Cells ◽

Cost Efficient ◽

Species Production

Development of clinical stem cell interventions are hampered by immature cell progeny under current protocols. Human mesenchymal stem cells (hMSCs) are characterized by their ability to self-renew and differentiate into multiple lineages. Generating hMSCs from pluripotent stem cells (iPSCs) is an attractive avenue for cost-efficient and scalable production of cellular material. In this study we generate mature osteoblasts from iPSCs using a stable expandable MSC intermediate, refining established protocols. We investigated the timeframe and phenotype of cells under osteogenic conditions as well as the effect of menaquinone-7 (MK-7) on differentiation. From day 2 we noted a significant increase in RUNX2 expression under osteogenic conditions with MK-7, as well as decreases in ROS species production, increased cellular migration and changes to dynamics of collagen deposition when compared to differentiated cells that were not treated with MK-7. At day 21 OsteoMK-7 increased alkaline phosphatase activity and collagen deposition, as well as downregulated RUNX2 expression, suggesting to a mature cellular phenotype. Throughout we note no changes to expression of osteocalcin suggesting a non-canonical function of MK-7 in osteoblast differentiation. Together our data provide further mechanistic insight between basic and clinical studies on extrahepatic activity of MK-7. Our findings show that MK-7 promotes osteoblast maturation thereby increasing osteogenic differentiation.

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Interleukin-17A inhibits adipocyte differentiation in human mesenchymal stem cells and regulates pro-inflammatory responses in adipocytes

Biochemical Pharmacology ◽

10.1016/j.bcp.2009.03.008 ◽

2009 ◽

Vol 77 (12) ◽

pp. 1835-1844 ◽

Cited By ~ 82

Author(s):

Jennifer H. Shin ◽

Dong Wook Shin ◽

Minsoo Noh

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipocyte Differentiation ◽

Inflammatory Responses ◽

Human Mesenchymal Stem Cells ◽

Interleukin 17A

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X-Ray Micro- and Nanodiffraction Imaging on Human Mesenchymal Stem Cells and Differentiated Cells

Biophysical Journal ◽

10.1016/j.bpj.2015.12.017 ◽

2016 ◽

Vol 110 (3) ◽

pp. 680-690 ◽

Cited By ~ 20

Author(s):

Marten Bernhardt ◽

Marius Priebe ◽

Markus Osterhoff ◽

Carina Wollnik ◽

Ana Diaz ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Human Mesenchymal Stem Cells ◽

X Ray ◽

Differentiated Cells

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P0-Related Protein Accelerates the Migration of Human Mesenchymal Stem Cells and Their Progeny by Modulating VLA-5 Interactions with Fibronectin.

Blood ◽

10.1182/blood.v108.11.1391.1391 ◽

2006 ◽

Vol 108 (11) ◽

pp. 1391-1391

Author(s):

Suzanne M. Watt ◽

Maria Roubelakis ◽

Sinead Forde ◽

Fengjuan Lu ◽

Grigorios Tsaknakis ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Inflammatory Responses ◽

Human Mesenchymal Stem Cells ◽

Leading Edge ◽

Cancer Surveillance ◽

Related Protein ◽

Transmembrane Receptors ◽

Type Iv ◽

First Time

Abstract Human mesenchymal stem cells (hMSCs) are capable of expansion, self renewal and generation of several tissues such as bone, fat and cartilage. They also generate microenviromental niche cells that regulate angiogenesis and hematopoiesis. During development and in the adult, hMSCs and their progeny migrate to specific niches where they provide support for hematopoietic cells or they play an important role in tissue regeneration, inflammatory responses and cancer surveillance. There is particular interest in understanding the molecular basis of hMSCs migration which involves numerous transmembrane receptors, cell adhesion and signaling molecules. Here, we demonstrate, for the first time, that human P0 - related protein, hPZR, has the ability to accelerate VLA-5-mediated migration of cultured hMSCs on the extracellular matrix (ECM) protein, fibronectin. This is mediated via the immunotyrosine-inhibitory motif (ITIM) sequences within the hPZR cytoplasmic domain, and the ECM ligand, since it cannot be recapitulated with the hPZR ITIM-less isoform, hPZRb, nor with other ECM components such as laminin or Type IV collagen. It is mediated by interactions with the docking phosphatase, SHP-2. The specificity of these results was confirmed by knocking-down hPZR and hPZRb using siRNA technology. We further demonstrated that hPZR clusters with VLA-5 at the leading edge of the cell, using high-resolution confocal microscopy together with immunoprecipitation and immunoblotting technologies. We propose that hPZR plays a key role in negatively regulating VLA-5 adhesion to fibronectin, a process critical for cell detachment from fibronectin and for the migration of hMSCs and their progeny.

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