A Real-Time PCR Approach to Evaluate Adipogenic Potential of Amniotic Fluid-Derived Human Mesenchymal Stem Cells

Gemmis, Paola de; Lapucci, Cristina; Bertelli, Matteo; Tognetto, Anna; Fanin, Erika; Vettor, Roberto; Pagano, Claudio; Pandolfo, Massimo; Fabbri, Andrea

doi:10.1089/scd.2006.15.719

Cited by 28 publications

(19 citation statements)

References 32 publications

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“…The amplification was performed with RT step (1 cycle at 50°C for 20 min) followed by initial activation of HotStar Taq DNA Polymerase at 95°C for 15 min and 40 cycles in three steps: 94°C for 10 s, 60°C for 15 s, 72°C for 30 s for C/EBP β, C/EBP δ, adipsin, PPARγ, UCP-1, vWF, KDR whereas for Flt-1 an additional step was added at 78°C for 2 s to analyze the fluorescence. The relative quantifications were performed by specific standard external curves as described [53] and the normalization was performed by parallel amplification of ribosomial 18S as described previously [54]. The specific oligo pairs for adipsin, PPARγ, UCP-1 and ribosomal 18S genes were already published [52], whereas the sequences of C/EBP β, C/EBP δ, vWF, Flt-1 and KDR were:…”

Section: Methodsmentioning

confidence: 99%

HIV-1 and recombinant gp120 affect the survival and differentiation of human vessel wall-derived mesenchymal stem cells

et al. 2011

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BackgroundHIV infection elicits the onset of a progressive immunodeficiency and also damages several other organs and tissues such as the CNS, kidney, heart, blood vessels, adipose tissue and bone. In particular, HIV infection has been related to an increased incidence of cardiovascular diseases and derangement in the structure of blood vessels in the absence of classical risk factors. The recent characterization of multipotent mesenchymal cells in the vascular wall, involved in regulating cellular homeostasis, suggests that these cells may be considered a target of HIV pathogenesis. This paper investigated the interaction between HIV-1 and vascular wall resident human mesenchymal stem cells (MSCs).ResultsMSCs were challenged with classical R5 and X4 HIV-1 laboratory strains demonstrating that these strains are able to enter and integrate their retro-transcribed proviral DNA in the host cell genome. Subsequent experiments indicated that HIV-1 strains and recombinant gp120 elicited a reliable increase in apoptosis in sub-confluent MSCs. Since vascular wall MSCs are multipotent cells that may be differentiated towards several cell lineages, we challenged HIV-1 strains and gp120 on MSCs differentiated to adipogenesis and endotheliogenesis. Our experiments showed that the adipogenesis is increased especially by upregulated PPARγ activity whereas the endothelial differentiation induced by VEGF treatment was impaired with a downregulation of endothelial markers such as vWF, Flt-1 and KDR expression. These viral effects in MSC survival and adipogenic or endothelial differentiation were tackled by CD4 blockade suggesting an important role of CD4/gp120 interaction in this context.ConclusionsThe HIV-related derangement of MSC survival and differentiation may suggest a direct role of HIV infection and gp120 in impaired vessel homeostasis and in genesis of vessel damage observed in HIV-infected patients.

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Section: Methodsmentioning

confidence: 99%

HIV-1 and recombinant gp120 affect the survival and differentiation of human vessel wall-derived mesenchymal stem cells

et al. 2011

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“…Although not necessarily with the aim to investigate pluripotency, inbetween a wide variety of different investigations reported differentiation of amniotic fluid-derived stem cells into neurogenic cells, or adipocytes, or smooth muscle cells, or chondrocytes, or osteoblasts [47,[60][61][62][63][64][65].…”

Section: Amniotic Fluid Stem Cells Display Multilineage Differentiatimentioning

confidence: 99%

Stem Cells in Amniotic Fluid as New Tools to Study Human Genetic Diseases

et al. 2007

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In future, the characterization and isolation of different human stem cells will allow the detailed molecular investigation of cell differentiation processes and the establishment of new therapeutic concepts for a wide variety of diseases. Since the first successful isolation and cultivation of human embryonic stem cells about 10 years ago, their usage for research and therapy has been constrained by complex ethical consideration as well as by the risk of malignant development of undifferentiated embryonic stem cells after transplantation into the patient's body. Adult stem cells are ethically acceptable and harbor a low risk of tumor development. However, their differentiation potential and their proliferative capacity are limited. About 4 years ago, the discovery of amniotic fluid stem cells, expressing Oct-4, a specific marker of pluripotent stem cells, and harboring a high proliferative capacity and multilineage differentiation potential, initiated a new and promising stem cell research field. In between, amniotic fluid stem cells have been demonstrated to harbor the potential to differentiate into cells of all three embryonic germlayers. These stem cells do not form tumors in vivo and do not raise the ethical concerns associated with human embryonic stem cells. Further investigations will reveal whether amniotic fluid stem cells really represent an intermediate cell type with advantages over both, adult stem cells and embryonic stem cells. The approach to generate clonal amniotic fluid stem cell lines as new tools to investigate molecular and cell biological consequences of human natural occurring disease causing mutations is discussed.

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“…Mesenchymal stem cells (MSCs) are the most commonly used cells in tissue engineering [2-4] and are found in several organic compartments, including the bone marrow, blood vessels, skin, and fat and muscle tissues [5]. MSCs can differentiate into osteogenic cells [6-8], chondrogenic cells [8,9], adipogenic cells [10] and cardiogenic cells [11] in response to different stimuli.…”

Section: Introductionmentioning

confidence: 99%

A new fibrin sealant as a three-dimensional scaffold candidate for mesenchymal stem cells

et al. 2014

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IntroductionThe optimization of an organic scaffold for specific types of applications and cells is vital to successful tissue engineering. In this study, we investigated the effects of a new fibrin sealant derived from snake venom as a scaffold for mesenchymal stem cells, to demonstrate the ability of cells to affect and detect the biological microenvironment.MethodsThe characterization of CD34, CD44 and CD90 expression on mesenchymal stem cells was performed by flow cytometry. In vitro growth and cell viability were evaluated by light and electron microscopy. Differentiation into osteogenic, adipogenic and chondrogenic lineages was induced.ResultsThe fibrin sealant did not affect cell adhesion, proliferation or differentiation and allowed the adherence and growth of mesenchymal stem cells on its surface. Hoechst 33342 and propidium iodide staining demonstrated the viability of mesenchymal stem cells in contact with the fibrin sealant and the ability of the biomaterial to maintain cell survival.ConclusionsThe new fibrin sealant is a three-dimensional scaffolding candidate that is capable of maintaining cell survival without interfering with differentiation, and might also be useful in drug delivery. Fibrin sealant has a low production cost, does not transmit infectious diseases from human blood and has properties of a suitable scaffold for stem cells because it permits the preparation of differentiated scaffolds that are suitable for every need.

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A Real-Time PCR Approach to Evaluate Adipogenic Potential of Amniotic Fluid-Derived Human Mesenchymal Stem Cells

Cited by 28 publications

References 32 publications

HIV-1 and recombinant gp120 affect the survival and differentiation of human vessel wall-derived mesenchymal stem cells

HIV-1 and recombinant gp120 affect the survival and differentiation of human vessel wall-derived mesenchymal stem cells

Stem Cells in Amniotic Fluid as New Tools to Study Human Genetic Diseases

A new fibrin sealant as a three-dimensional scaffold candidate for mesenchymal stem cells

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