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5. MMP-2, TIMP-2, TAZ and MEF2a Transcript Expression in Osteogenic and Adipogenic Differentiation of Porcine Mesenchymal Stem Cells

). Osteoblastogenic effects of dexamethasone through upregulation of TAZexpression in rat mesenchymal stem cells. J. Steroid. Biochem. Mol. Biol., 116: 86-92. Hoshiba T., Kawazoe N., Chen G. (2012). The balance of osteogenic and adipogenic differentiation in human mesenchymal stem cells by matrices that mimic stepwise tissue development. Biomaterials, 33: 2025-2031. Hu E., Tontonoz P., Spiegelman B.M. (1995). Transdifferentiation of myoblasts by the adipogenic transcription factors PPARγand C/EBPα. Proc. Nat. Acad. Sci. USA., 92: 9856- -9860

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Isolation and cellular properties of mesenchymal stem cells from human periosteum

Abstract

Background: Cell-based therapy has achieved good functional recovery for tissue repair. Mesenchymal stem cells (MSCs) exhibit multilineage potential, long-term viability, and capacity for self-renewal. Periosteum-derived mesenchymal stem cells (PD cells) may be an attractive cell source for tissue engineering because of their easy accessibility and reduced ethical concerns.

Objectives: To isolate and investigate the phenotypic and functional characteristics of mesenchymal stem cells derived from human periosteum. We also examined the differentiation of PD cells with a trilineage differentiation assay to determine whether they were MSCs.

Materials and Methods: Periosteum-derived cells were cultured in osteogenic, chondrogenic or adipogenic media to evaluate their multilineage differentiation potential. Adherent fibroblast-like cells were analyzed by flow cytometry for MSC cell surface markers. Differentiation of PD cells into osteogenic, chondrogenic, and adipogenic lineages was also evaluated by von Kossa, Alizarin red, Alcian blue, and oil red O stains, respectively. Expression of mesenchymal stem cell markers were assessed using reverse transcriptase-polymerase chain reaction (RT-PCR) analysis.

Results: We successfully isolated and expanded MSCs from human periosteum. Flow cytometry revealed that PD cells were positive for mesenchymal adhesion cell markers (CD29, CD44, CD90, and CD105) and negative for hematopoietic markers (CD34 and CD45). In osteogenic differentiation, calcium accumulation (positive von Kossa and Alizarin red) and RUNX2, alkaline phosphatase, collagen type I, osteopontin genes were detected. In adipogenic differentiation, the cells displayed oil red O positive and expressed lipoprotein lipase and peroxisome proliferator-activated receptor-gamma (PPAR-γ) associated with adipogenesis. The cells grown in chondrogenic conditions were positively stained for Alcian blue and expressed SOX-9.

Conclusion: PD cells presented osteogenic, chondrogenic, and adipogenic differentiation abilities in vitro and could provide an alternative cellular source for tissue repair in clinical applications.

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The Quality of Porcine Mesenchymal Stem Cells and Their Osteo- and Adipogenic Cell Derivatives – The Level of Proapoptotic Bad Protein Expression / Jakość Mezenchymalnych Komórek Macierzystych Świni Oraz Ich Pochodnych Zróżnicowanych W Kierunku Komórek Szeregu Osteo- I Adipogennego – Poziom Ekspresji Proapoptotycznego Białka Bad

Abstract

The aim of the research was to evaluate the quality of porcine mesenchymal stem cells (MSCs) and MSC-derived osteoblasts/osteocytes (bone cells) and adipocytes (fat cells). This evaluation was performed on the basis of molecular analysis for proapoptotic BAD protein expression. MSCs isolated from the pig bone marrow were cultured in vitro for five weeks in three types of culture media: differentiating towards the osteoblasts/osteocytes (O) and adipocytes (A) and non-differentiating, control medium (C). In all groups of cells, the relative extent of BAD protein expression was estimated by western blotting. Significant differences in the posttranslational abundance of BAD proteins were noted between MSCs differentiated into the osteogenic and the adipogenic cell lineages (P<0.05). Summarizing the results, we conclude that posttranslational level of BAD protein expression can be used as a reliable marker for assessing the quality of both MSCs and their cell derivatives. Interestingly, the semi-quantitative profile of BAD protein expression in differentiated cells turned out to be lower than that observed in undifferentiated cells, demonstrating that the culture conditions used for pro-osteogenic or pro-adipogenic cellular transformation did not affect negatively the quality of MSCs.

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The Antidepressants and the Metabolic Syndrome

Syndrome: Systematic Review and Meta- Analysis Study. 2016;18. 64. Sun BK, Kim JH, Choi J-S, Hwang S-J, Sung J-H. Fluoxetine Decreases the Proliferation and Adipogenic Differentiation of Human Adipose-Derived Stem Cells. Int. J. Mol. Sci. Multidisciplinary Digital Publishing Institute (MDPI); 2015;16:16655–68. 65. Cloonan SM, Williams DC. The antidepressants maprotiline and fluoxetine induce Type II autophagic cell death in drug-resistant Burkitt’s lymphoma. Int. J. Cancer. 2011;128:1712–23. 66. Li H, Fong CC, Chen Y, Cai G, Yang M. Imipramine inhibits

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Proliferation And Differentiation Potential Of Canine Synovial Fluid Cells

Abstract

The aim of this study was to determine whether synovial fluid (SF) of dogs contains cells that have characteristics of MSCs and to describe their differentiation potential. SF adherent cells from 5 young German shepherd dogs (average 3.8 ± 0.9 years) were expanded (37°C, 5% CO2, humidified atmosphere) three weeks before their phenotype was characterized by flow-cytometry for the presence of CD90 and CD34. Population doubling time (PDT), number of CFU-F and adipogenic, osteogenic and chondrogenic potentials have been determined in vitro. In early passages PTD was 31 ± 10 hours and expansion fold after 3 sub cultivations (9 days) theoretically could be 372 ± 134. At P1, 0.55 ± 0.05% of SF cells had the ability to form CFU-F. Sixty-six percent of cells expressed CD90 and none of the cells expressed markers of hematopoietic cells. Oil Red O staining has shown accumulation of fat droplets in cells grown in adipogenic medium, while deposits of calcium in the osteogenic medium were evidenced with Alizarin red staining. SF cultured in hondrogenic and control medium in three-dimensional conditions formed a cartilage-like tissue. Alcian blue staining of pellets’ slides have shown a significant amount of glycosaminoglycans (GAGs) and immunohistochemistry analysis documented collagen type II expression. The amount of GAGs in pellets grown in both conditions showed no difference. SF cells in vitro exhibited osteogenic, adipogenic and chondrogenic differentiation potentials, suggesting the presence of different mesenchymal progenitors. These results also demonstrated that SF cells have a spontaneous chondrogenic potential that should be further explored for possible tissue engineering protocols.

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Isolation of Dental Pulp Stem Cells and their In Vitro Differentiation into Odontoblast-like Cells

Isolation of Dental Pulp Stem Cells and their In Vitro Differentiation into Odontoblast-like Cells

Background: Recently, tooth tissue engineering has attracted more and more attention. Stem cell based tissue engineering is thought to be a promising way to replace the missing tooth. Mesenchymal stem cells (MSCs) are multipotent stem cells which can differentiate into a variety of cell types.

Material and Methods: We isolated stem cells from dental pulp and measured their self-renewal capacities. Adipogenic, chondrogenic as well as odontogenic differentiation potentials were investigated, using bone morphogenic protein 2 (BMP2) for the odontogenic differentiation.

Results: The cumulative number of the isolated cells was high. Polycomb ring finger oncogene (Bmi1) and Signal transducer and activator of transcription 3 (Stat3) were continuously expressed suggesting longer proliferative lifespan and self-renewal capacity of the isolated cells. Peroxisome proliferatoractivated receptor was expressed showing adipogenic conversion that was also confirmed by positive staining of cells with Oil red O stain and chondrogenic differentiation was confirmed by positive staining of cells with Alcian blue stain. BMP2 stimulated the expression of dentin sialophosphoprotein (DSPP) and enamelysin, indicating successful odontogenic differentiation that was also confirmed by the positive staining of the cells with Alizarin red stain.

Conclusion: Thus, adult pulp contains stem cells, which are useful for cell therapy with BMP2 for dentin regeneration.

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Characterisation and In Vivo Safety of Canine Adipose-Derived Stem Cells

Abstract

The study characterises canine adipose-derived stem cells (cASCs) in comparison to human ASCs (hASCs) and tests their safety in a canine model after intravenous administration. cASCs from two dogs were cultured under hypoxic conditions in a medium supplemented with autologous serum. They were plastic adherent, spindle-shaped cells that expressed CD73, CD90, and CD44 but lacked CD45, CD14, HLA-DR, and CD34. cASCs differentiated toward adipogenic, osteogenic, and chondrogenic lineages, although adipogenic differentiation capacity was low. Blast transformation reaction demonstrated that these cells significantly suppress T-cell proliferation, and this ability is dose-dependent. Intravenous administration of a cell freezing medium, therapeutic dose of cASCs (2 × 106 live cells/kg), and five times higher dose of cASCs showed no significant side effects in two dogs. Microscopic tissue lesions were limited to only mild, non-specific changes. There were no signs of malignancy. The results of the study indicate that cASCs are similar to hASCs and are safe for therapeutic applications in a canine model. The proposed methodology for ASC preparation on a non-routine basis, which includes individually optimised cell culture conditions and offers risk-adapted treatment, could be used for future personalised off-the-shelf therapies, for example, in myocardial infarction or stroke.

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Allogeneic Bone Marrow Multipotent Mesenchymal Stromal Cells and Polytrauma Repair: The Role of Fractionated on the Basis of Molecular Mass Red Beetroot Juice in the Prevention of Transplanted Cells Side Effects in Rats

Red beetroot (Beta vulgaris) juice (RBJ) is used as a traditional medicine for treatment of anemia. It has been shown that beetroot juice decreases blood pressure, provides a protective effect on blood vessels and has antioxidant and anticancerogenic properties. In the case of polytrauma it might have beneficial effects because of its antioxidant and anti-inflammatory properties as well as antimicrobial activity. It is also well-known that RBR juice can induce undesirable side effects, e.g. flatulent stomach, nausea and other unpleasant reactions. Therefore, it seems prospective to develop red beetroot juice based on its natural compound composition free of undesirable side effects, which could then be used in combination with bone marrow multipotent mesenchymal stromal cells (BM MMSC) transplantation in the case of polytrauma. The aim of the study was to evaluate the therapeutic effect of allogeneic BM MMSC transplantation in rats with experimental polytrauma and to analyse red beetroot fractions separated on the basis of molecular weight in regard to their ingestion impact on cell transplantation efficacy. Red beet juice was fractionated by ultrafiltration (cut-off-point 20 kDa). Total phenolic compound concentration in the final product practically did not decrease. The product was tested in vitro and in vivo. Unlike native juice, fractionated RBJ in vitro suppressed BM MMSC adipogenic (60-71%, P < 0.05) and stimulated osteogenic differentiation (124%, P < 0.05). Experimental polytrauma in rats was modelled by causing three fractures and haemorrhagic shock. Animals were randomised in five groups: 1) normal control; 2) polytrauma; 3) polytrauma + i/v BM MMSC transplantation 36 h and 5 days after surgery; 4) polytrauma + fractionated RBJ administration per os 1ml/d, and 5) polytrauma + BM MMSCs + fractionated RBJ. Transplantation of allogeneic BM MMSCs in rats with experimental polytrauma stimulated bone fracture reparation, but caused plethora in viscera and dystrophic changes in lungs. Combination of BM MMSCs and fractionated RBJ resulted in better bone reparation and significant hematopoiesis stimulation.

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Chronic fluoxetine treatment induces lipid accumulation but does not alter the expression of Pref-1 in rat adipose tissue

serotonin reuptake inhibitors on three sex steroids in two versions of the aromatase enzyme inhibition assay and in the H295R cell assay, Toxicol. in Vitro 29 (2015) 1729-1735; https://doi.org/10.1016/j.tiv.2015.07.005 8. B. K. Sun, J. H. Kim, J. S. Choi, S. J. Hwang and J. H. Sung, Fluoxetine decreases the proliferation and adipogenic differentiation of human adipose-derived stem cells, Int. J. Mol. Sci. 16 (2015) 16655-16668; https://doi.org/10.3390/ijms160716655 9. M. Kolcsár, Z. Gáll, I. L. Bíró, L. I. Bába, A. Imre, M. T. Dogaru and O

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Review of the role of basic fibroblast growth factor in dental tissue-derived mesenchymal stem cells

]. Similarly, the effects of bFGF on adipogenic differentiation are controversial. These contradictory results may be a consequence of different cell types, concentrations, exposure times, and culture conditions. DMSCs have been introduced as a stem cell source because of their accessibility and availability. MSCs can be isolated from various dental-related tissues, including dental pulp, periodontal ligaments, apical papilla, and dental follicles [ 14 ]. The isolated cells exhibit the stem cell characteristics, including the expression of mesenchymal stem cell markers and

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