Sant'Ambrogio di Torino, Italy
Sant'Ambrogio di Torino, Italy

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Moggio A.,Center for Molecular Biotechnology | D'Armento G.,University of Turin | Bussolati B.,Center for Molecular Biotechnology
Organogenesis | Year: 2012

Different approaches for the isolation of stem/progenitor cells have been reported, including stem cell selection in stringent culture conditions. We evaluated the possibility of isolating human progenitor cells from surgical specimens preserved by under vacuum sealing and cooling, a clinical practice approached by several hospitals as alternative to formalin. Renal tissue samples (n = 20) maintained under vacuum from 6 to 48 h at 4°C were used to isolate human renal CD133+ progenitor cells. We obtained CD133+ progenitors from unsorted cells derived from disaggregated tissues from each sample. Phenotypic characterization as well as in vitro and in vivo differentiation of the obtained CD133+ lines showed results comparable with sorted CD133+ cells obtained from fresh tissue. These results indicate that the process of sealing under vacuum and cooling appears as a suitable tissue treatment to isolate hypoxia resistant cells, such as human stem/progenitor cells, and that this procedure can be exploited to render the extraction of stem cells from human samples more practical and feasible. © 2012 Landes Bioscience.


Tetta C.,Fresenius Medical Care Deutschland GmbH | Bruno S.,Sis Ter | Bruno S.,Center for Molecular Biotechnology | Fonsato V.,Center for Molecular Biotechnology | And 2 more authors.
Organogenesis | Year: 2011

Microvesicles (MVs) are released by almost all cells in resting and activated conditions. First described several years ago, it is only recently that their mechanisms of actions are being elucidated, and their potential role in health and disease is drawing increasing attention. The main function of MVs is the signaling through specific interactions with target cells and the transferring of gene products. Gaining further insights into the molecular specificity of MVs has allowed identifying the cellular source and may provide new diagnostic tools in the future. Indeed, an increasing body of evidence indicates that MVs are capable of mediating tissue repair in models of acute kidney and liver injury.this review, we will discuss the mechanisms through which MVs from stem cells may act on target cells and may modify the response to injury. Furthermore, MVs from inflammatory cells are suspected to be involved in various diseases such as, cardiovascular and renal diseases, pathological pregnancy, tumors and sepsis. MVs are no doubt also involved in modulating immunity and future studies will clarify their functional role in negatively modulating the cell response. Their role in physiological and pathological processes is increasingly appreciated. Depending on the cell source and the condition, MVs may be either beneficial or detrimental to the host. The recognition of their pathogenetic role may suggest new approaches to future therapies. © 2011 Landes Bioscience.


Camussi G.,Center for Molecular Biotechnology | Deregibus M.C.,Center for Molecular Biotechnology | Bruno S.,Sis Ter | Cantaluppi V.,Center for Molecular Biotechnology | Biancone L.,Center for Molecular Biotechnology
Kidney International | Year: 2010

Microvesicles (MVs) are circular fragments of membrane released from the endosomal compartment as exosomes or shed from the surface membranes of most cell types. An increasing body of evidence indicates that they play a pivotal role in cell-to-cell communication. Indeed, they may directly stimulate target cells by receptor-mediated interactions or may transfer from the cell of origin to various bioactive molecules including membrane receptors, proteins, mRNAs, microRNAs, and organelles. In this review we discuss the pleiotropic biologic effects of MVs that are relevant for communication among cells in physiological and pathological conditions. In particular, we discuss their potential involvement in inflammation, renal disease, and tumor progression, and the evidence supporting a bidirectional exchange of genetic information between stem and injured cells. The transfer of gene products from injured cells may explain stem cell functional and phenotypic changes without the need of transdifferentiation into tissue cells. On the other hand, transfer of gene products from stem cells may reprogram injured cells to repair damaged tissues. © 2010 International Society of Nephrology.


Camussi G.,Center for Molecular Biotechnology | Cantaluppi V.,Center for Molecular Biotechnology | Deregibus M.C.,Center for Molecular Biotechnology | Gatti E.,International Research and Development | And 3 more authors.
Contributions to Nephrology | Year: 2011

The main function of microvesicles (MVs) is signaling through specific interactions with target cells and transferring gene products. Therefore, they may participate in physiological and pathological processes. Gaining further insights into the molecular specificity of MVs has allowed identifying the cellular source and may provide new diagnostic tools in the future. Indeed, an increasing body of evidence indicates that MVs may offer prognostic information in various diseases such as chronic inflammation, cardiovascular and renal diseases, pathological pregnancy, tumors, and sepsis. The presence of MVs in body fluids makes them readily accessible. Their number, cellular origin, composition and function can be dependent on the state of the disease. In sepsis for example, activated endothelial cells may shed MVs that might trigger leukocyte and monocyte production and release pro-oxidant and inflammatory mediators. MVs from platelets may trigger disseminated intravascular coagulopathy. MVs are no doubt also involved in modulating immunity and future studies will clarify their functional role in negatively modulating the cell response. In addition, the recognition of the signals delivered by MVs may open new therapeutic strategies. The removal of harmful MVs from plasma may be beneficial in pathological conditions where MVs deliver thrombogenic and inflammatory signals. On the other hand, MVs derived from stem cells may reprogram altered functions in target cells suggesting that they could be exploited in regenerative medicine to repair damaged tissues. We will discuss the role of stem cell-derived MVs in the repair of acute kidney injury. Copyright © 2011 S. Karger AG, Basel.


Bussolati B.,Center for Molecular Biotechnology | Deregibus M.C.,Center for Molecular Biotechnology | Camussi G.,Center for Molecular Biotechnology
Current Vascular Pharmacology | Year: 2010

The neo-angiogenesis process is crucial for solid tumor growth and invasion, as the vasculature provides metabolic support and access to the circulation. Tumor blood vessels differ from normal vessels by altered morphology, blood flow and permeability, and the "switch" of endothelial cells to an angiogenic phenotype is considered a hallmark of the malignant process. Recent evidence indicates that tumor-derived endothelial cells (TEC) possess a distinct and unique phenotype differing from normal endothelial cells at the molecular and functional levels. The anti-angiogenic therapies developed to date are based on tumor endothelial cells being genetically normal. However, it has recently been shown that TEC derived from different tumors are genetically unstable and may acquire resistance to drugs. It has been suggested that TEC may acquire cytogenetic abnormalities within the tumor microenvironment. We found that TEC from different tumors share characteristics in terms of pro-angiogenic properties, survival and resistance to chemotherapy in respect to non-tumor endothelial cells and maintain in vitro an immature pro-angiogenic phenotype in the absence of tumor cells. This was associated with an up-regulation of the AKT/PI3K pathway, involved in the repression of the anti-angiogenic factors thrombospondin-1 and PTEN, and the presence in TEC of the embryonic transcription factor PAX2, responsible for the expression of immature endothelial markers such as NCAM. The in vivo inhibitions of these pathways were shown to display an anti-angiogenic effect on TEC. This review considers the current studies on TEC abnormalities and discusses the hypothesis that at least part of tumor vessels may derive from an intra tumor ongoing embryonic-like vasculogenesis or from tissue endothelial cells switched to angiogenesis from genetic information transmitted from the tumor. © 2010 Bentham Science Publishers Ltd.


PubMed | Center for Molecular Biotechnology
Type: Journal Article | Journal: Organogenesis | Year: 2013

Different approaches for the isolation of stem/progenitor cells have been reported, including stem cell selection in stringent culture conditions. We evaluated the possibility of isolating human progenitor cells from surgical specimens preserved by under vacuum sealing and cooling, a clinical practice approached by several hospitals as alternative to formalin. Renal tissue samples (n = 20) maintained under vacuum from 6 to 48 h at 4C were used to isolate human renal CD133(+) progenitor cells. We obtained CD133(+) progenitors from unsorted cells derived from disaggregated tissues from each sample. Phenotypic characterization as well as in vitro and in vivo differentiation of the obtained CD133(+) lines showed results comparable with sorted CD133(+) cells obtained from fresh tissue. These results indicate that the process of sealing under vacuum and cooling appears as a suitable tissue treatment to isolate hypoxia resistant cells, such as human stem/progenitor cells, and that this procedure can be exploited to render the extraction of stem cells from human samples more practical and feasible.


PubMed | Center for Molecular Biotechnology
Type: Journal Article | Journal: Histology and histopathology | Year: 2010

It has been commonly supposed that adult stem cells co-localize with supporting cells within specific regions or specialized microenvironment in each tissue/organ, called stem cell niche. This concept was based on the assumption that stem cells are intrinsically hierarchical in nature. However, recent data indicate that stem cells may represent a continuum with reversible alterations in phenotype taking place during the transit through cell cycle. Based on this dynamic interpretation it has been suggested that the so-called niche is represented by a single or only few cell types continually adjusting their phenotype and function to individual circumstances. A critical component in the regulation of the continuum of stem cell phenotypes is the microenvironment. In this context, microvesicles (MVs) account for the transfer of genetic information between cells. Originally considered inert cellular debris, MVs are increasingly recognized to be important mediators of cell-to-cell communication. MVs may transfer receptors, proteins, mRNA and microRNA to target cells via specific receptor-mediated interaction. In stem cell biology the exchange of genetic information may be bidirectional from stromal to stem cells. In the context of tissue injury the MV-mediated transfer of genetic information may reprogram the phenotype of stem cells to acquire features of the injured tissue cells. In addition, MVs derived from stem cells may induce de-differentiation of cells which have survived injury with a cell cycle re-entry that may allow tissue regeneration. In the present review we discuss the possibility of a continuous genetic modulation of stem cells by a MV-mediated transfer of information between cells.

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