Mesoangioblasti di topo producono e rilasciano nello spazio extracellulare vescicole di membrana contenenti MMP9 e FGF2 in forma attiva

Positive or negative involvement of heat-shock proteins in multiple sclerosis pathogenesis: an overview.

Shedding of membrane vesicles containing HSP70 and FGF-2 from A6 stem cells.

Sistemas de endomembranas (RE y GOLGI), sistema vacuolar y lisosomas

Mouse A6 stem cells release active FGF-2 into extracellular space through plasma membrane vesicles

In this study, mouse mesoangioblasts were seeded onto bidimensional matrices within three-dimensional porous scaffolds of poly (L-lactic acid) (PLLA), in the presence or absence of a type I collagen coating. The cells were observed under a scanning electron microscope and tested for their adhesion, survival and proliferation. Immunolocalization of heat shock protein (Hsp) 70, an abundant and ubiquitous intracellular protein in these cells, was also performed in sectioned cell-containing scaffolds under a confocal fluorescence microscope to determine if in situ analysis of intracellular constituents was feasible. The data show that PLLA films allow direct cell adhesion and represent an optim…

Nichel,piombo e cadmio inducono risposte cellulari differenti, attivando la sintesi di differenti HSP70 in embrioni di ricci di mare

Selezione di cloni più resistenti allo stress ossidativo dalla popolazione di cellule staminali mesoangioblasti

Heat-Shock Proteins in Autoimmunity

Heat shock proteins (HSPs), also known as “stress proteins,” are among the highly conserved and immunogenic proteins shared among diverse groups of microbial agents and mammals [1]. Heat and other types of stressful stimuli can increase the cellular expression of HSPs. These proteins have been categorized into different families according to their molecular mass, for example, HSP110, HSP90, HSP70, HSP60, HSP40, HSP20-30, and HSP10 [1–3]. For uniformity, guidelines for the nomenclature of various human HSP families have been proposed [4]. Under physiological conditions, the ubiquitously distributed HSPs maintain the integrity and function of other cellular proteins in stressful conditions. H…

EXTRACELLULAR VESICLES IN CEREBROSPINAL FLUID AS MARKERS OF CNS INFLAMMATION

Mouse mesoangioblast behaviour when subjected to cellular stress

Vanadium perturbs the fertilization outcome and the metalloproteinase activity in sea urchin embryos

Metal toxicology represents a current major topic due to the disper- sion of these elements in the environment. Metals are released from both natural sources and industrial activities. Some of them have also a clinical interest due to their application as metallodrugs (i.e., Pt, Cu, Au, Ru, and Y) or in medical diagnosis (Gd).1,2 Recently, V derivatives are considered as potential therapeutic factors in some diseases (e.g., obesity, diabetes, cancer, neurodegenerative and heart disorders). As a consequence, pharmaceutical residues could repre- sent emerging pollutants of aquatic environments, as wastewater treatment plants do not sufficiently remove these compounds3. Embryonic models repres…

Membrane vesicles containing matrix metalloproteinase-9 and fibroblast growth factor-2 are released into the extracellular space from mouse mesoangioblast Sstem cells

Certain proteins, including fibroblast growth factor-2 (FGF-2) and matrix metalloproteinase-9 (MMP-9), have proved very effective in increasing the efficacy of mesoangioblast stem cell therapy in repairing damaged tissue. We provide the first evidence that mouse mesoangioblast stem cells release FGF-2 and MMP-9 in their active form through the production of membrane vesicles. These vesicles are produced and turned over continuously, but are stable for some time in the extracellular milieu. Mesoangioblasts shed membrane vesicles even under oxygen tensions that are lower than those typically used for cell culture and more like those of mouse tissues. These findings suggest that mesoangioblast…

Extracellular Vesicles: The New Frontier of Stem Cell Regenerative Medicine?

Regenerative medicine aims to repair damaged or missing cells, tissues or organs for the treatment of various diseases, poorly managed with conventional drugs and medical procedures. To date there are different approaches to obtain these results. Multimodal regenerative methods include transplant of healthy organs, tissues, or cells, body stimulation to activate a self healing response in damaged tissues, as well as the combined use of cells and bio-degradable scaffold to obtain functional tissues. Certainly, stem cells and derived products are promising tools in regenerative medicine due to their ability to induce de novo tissue formation and/or promote tissue and organ repair and regenera…

Cellule staminali A6 di topo producono vescicole che contengono HSP70 e FGF-2

Autocrine role of extracellular Hsp70 in mesoangioblast migration capability

MMP2 synthesis in mouse mesoangioblast stem cells is highly regulated

Metalloproteasi nella biologia dei mesoangioblasti di topo

Gastrulation in sea urchin and amphibian embryos

Espressione basale dell’HSP70 inducibile in differenti linee staminali di topo.

Another function of Hsp70 in mesoangioblast stem cells

In recent years, it has been demonstrated that Hsp70 is released in the extracellular space under normal cell culture conditions, and this release is mediated through exosomes. We have demonstrated that A6 cells, a clone of mouse mesoangioblasts, produce and release in the extracellular space membrane vesicles, independently of culture growth conditions. These vesicles contains both structural proteins and biological active molecules, such as FGF-2 and the metalloproteinases MMP 2 and 9. We have also demonstrated that A6 vesicles contain HspP70 and its release is highly regulated. Some of the intracellular Hsp70 is localized on lipid rafts and its concentration in insoluble fraction increas…

Cell clone selected from mab stem cell is more resistent to oxidative stress and retain some ability that could be useful for cell therapy.

Extracellular release of Hsp70 from A6 mouse stem cells

Paracrine roles of extracellular vesicles released by mouse mesoangioblasts

Extracellular vesicles (EV) represent an important mediator of cell-to-cell communication and are involved in both autocrine and paracrine signaling, with a critical role in a number of physiological and pathological conditions.1 The bioactive molecules contained within EV simultaneously activate several different pathways resulting in the synergistic stimulation of target cells. The discovery and characterization of EV have added a novel understanding to regenerative medicine, namely the finding that stem cells are an abundant source of EV.1-2 A6 mouse mesoangioblasts, vessel-associated multipotent progenitor stem cells that are capable of differentiating into different mesodermal cell typ…

Vanadium Toxicity Monitored by Fertilization Outcomes and Metal Related Proteolytic Activities in Paracentrotus lividus Embryos

Metal pharmaceutical residues often represent emerging toxic pollutants of the aquatic environment, as wastewater treatment plants do not sufficiently remove these compounds. Recently, vanadium (V) derivatives have been considered as potential therapeutic factors in several diseases, however, only limited information is available about their impact on aquatic environments. This study used sea urchin embryos (Paracentrotus lividus) to test V toxicity, as it is known they are sensitive to V doses from environmentally relevant to very cytotoxic levels (50 nM; 100 nM; 500 nM; 1 µM; 50 µM; 100 µM; 500 µM; and 1 mM). We used two approaches: The fertilization test (FT) and …

Mechanisms of Ca2+ liberation at fertilization

The mechanisms underlying the Ca2+ release at fertilization of several animal organisms are reported. Four main classical theories are described, i.e., that of Ca2+ release following simple sperm contact and a G protein stimulation; that of simple sperm contact followed by a tyrosine kinase receptor activation; that of the necessity of introduction by sperm into the egg of molecules for Ca2+ release; and that the molecule introduced into the marine eggs for Ca2+ release is the same Ca2+. Two other mechanisms for Ca2+ release are also illustrated: that of ryanodine receptor stimulation and that of NAADP formation.

Nickel, lead, and cadmium induce differential cellular responses in sea urchin embryos by activating the synthesis of different HSP70s.

Treatment with heavy metals, such as nickel, lead or cadmium, elicits different cellular stress responses according to the metal used and the length of treatment. In Paracentrotus lividus embryos the inducible forms of HSP70 (HSP70/72) are different in molecular mass from the constitutively expressed HSP75, and they can be used as markers of cellular stress. Even a short treatment with each metal induces the synthesis of HSP70/72 which remain stable for at least 20 h and differ little in their isoelectric points. Continuous treatment from fertilization with nickel or lead produces late irregular pluteus embryos, with peak HSP70/72 synthesis at blastula followed by the arrest of synthesis by…

Constitutive hsp70 is essential to mitosis during early cleavage of Paracentrotus lividus embryos: The blockage of constitutive hsp70 impairs mitosis

Localization of constitutive hsp70 in eggs and early embryos of sea urchin Paracentrotus lividus is shown by means of in situ immunostaining. An accumulation of this protein is shown in the mitotic structures (asters, spindles and centrosomes). Microinjection of anti-hsp70 antibodies into eggs causes impairment of formation of mitotic structures and of cell division. This impairment goes from a complete mitotic block, to irregular mitotic apparatus formation with irregular cleavage, depending upon the antibody concentration. The localization of hsp70 after antibody microinjection is also described. Blockage of mitotic apparatus formation by nocodazole also blocks the concentration of hsp70 …

Prognostic and Functional Significant of Heat Shock Proteins (HSPs) in Breast Cancer Unveiled by Multi-Omics Approaches

Simple Summary In this study, we investigated the expression pattern and prognostic significance of the heat shock proteins (HSPs) family members in breast cancer (BC) by using several bioinformatics tools and proteomics investigations. Our results demonstrated that, collectively, HSPs were deregulated in BC, acting as both oncogene and onco-suppressor genes. In particular, two different HSP-clusters were significantly associated with a poor or good prognosis. Interestingly, the HSPs deregulation impacted gene expression and miRNAs regulation that, in turn, affected important biological pathways involved in cell cycle, DNA replication, and receptors-mediated signaling. Finally, the proteomi…

Identification of an HSF2-like factor in sea urchin embryos and its localization in primary mesenchime cells

Intracellular and extracellular Hsp70 in A6 mouse stem cells

Oxidative stress preconditioning of mouse perivascular myogenic progenitors selects a subpopulation of cells with a distinct survival advantage in vitro and in vivo

AbstractCell engraftment, survival and integration during transplantation procedures represent the crux of cell-based therapies. Thus, there have been many studies focused on improving cell viability upon implantation. We used severe oxidative stress to select for a mouse mesoangioblast subpopulation in vitro and found that this subpopulation retained self-renewal and myogenic differentiation capacities while notably enhancing cell survival, proliferation and migration relative to unselected cells. Additionally, this subpopulation of cells presented different resistance and recovery properties upon oxidative stress treatment, demonstrating select advantages over parental mesoangioblasts in …

Hsp70 and Its Molecular Role in Nervous System Diseases

Heat shock proteins (HSPs) are induced in response to many injuries including stroke, neurodegenerative disease, epilepsy, and trauma. The overexpression of one HSP in particular, Hsp70, serves a protective role in several different models of nervous system injury, but has also been linked to a deleterious role in some diseases. Hsp70 functions as a chaperone and protects neurons from protein aggregation and toxicity (Parkinson disease, Alzheimer disease, polyglutamine diseases, and amyotrophic lateral sclerosis), protects cells from apoptosis (Parkinson disease), is a stress marker (temporal lobe epilepsy), protects cells from inflammation (cerebral ischemic injury), has an adjuvant role i…

Ectosomes containing HSP70 and FGF-2 are released from mouse A6 stem cells

Sea urchin HSF activity in vitro and in transgenic embryos.

Evidence is provided for the presence at the physiological temperature of 20 degrees C of a heat shock transcriptor factor, HSF, in the nuclei of P.lividus embryos. This HSF is able to specifically bind in vitro the heat shock element, HSE, of the promoter of the hsp70 gene i.v., as suggested by DNA-protein binding reactions and DNAse I protection assays. Upon heat-shock, at the temperature of 31 degrees C, its ability to bind the HSE units becomes much higher. The HSF activated by heat-shock drives in vivo the transcription of the beta-galactosidase reporter gene in transgenic sea urchin gastrulae. An ATF-like transcription factor, widely described in other organisms but not at all in sea …

Localization of HSP70, Cdc2, and cyclin B in sea urchin oocytes in non-stressed conditions.

In Paracentrotus lividus embryos, a Mediterranean sea urchin species, HSP70 is present in all the cells. During cell division it localizes under normal growth conditions on the centrosomes and on the whole isolated mitotic apparatus. Now, in situ hybridization, Western blot analyses, and immunohistochemistry show that the HSP70 mRNA is present in both small and large P. lividus oocytes, that all four isoforms of HSP70 can be found also in the oocytes, and that a certain amount of HSP70 localizes on asters and spindles during polar body formation. Moreover, two representative cell-cycle related proteins, cyclin B, and Cdc2, are present both in small and large oocytes, concentrating in the ge…

Hsp70 functions: inside and outside the cell

Differentiation capability of clones of mouse mesoangioblast stem cells resistant to oxidative stress.

Diversification of spatiotemporal expression and copy number variation of the echinoid hbox12/pmar1/micro1 multigene family

Changes occurring during evolution in the cis-regulatory landscapes of individual members of multigene families might impart diversification in their spatiotemporal expression and function. The archetypal member of the echinoid hbox12/pmar1/micro1 family is hbox12-a, a homeobox-containing gene expressed exclusively by dorsal blastomeres, where it governs the dorsal/ventral gene regulatory network during embryogenesis of the sea urchin Paracentrotus lividus. Here we describe the inventory of the hbox12/pmar1/micro1 genes in P. lividus, highlighting that gene copy number variation occurs across individual sea urchins of the same species. We show that the various hbox12/pmar1/micro1 genes grou…

Intracellular and extracellular Hsp70 in mouse mesoangioblast stem cells

Stress response and apoptosis in mesoangioblasts stem cells

Starfish and Xenopus oocyte maturation

Treatment with heavy metals, such as nickel, lead or cadmium, elicits different cellular stress responses according to the metal used and the length of treatment. In Paracentrotus lividus embryos the inducible forms of HSP70 (HSP70/72) are different in molecular mass from the constitutively expressed HSP75, and they can be used as markers of cellular stress. Even a short treatment with each metal induces the synthesis of HSP70/72 which remain stable for at least 20h and differ little in their isoelectric points. Continuous treatment from fertilization with nickel or lead produces late irregular pluteus embryos, with peak HSP70/72 synthesis at blastula followed by the arrest of synthesis by …

Hsp70 and NF-kB are involved in mouse mesoangioblast stem cells migration.

Stress response in mouse stem cells

Hsp70 level regulates MMP2 expression in mesoangioblast stemj cells

Mesoangioblast stem cell population is non-omogeneous as revealed by transcriptome analysis after a severe oxidative stress.

Esocitosi di Hsp70 mediante vescicole in cellule staminali di topo

Heat Shock Proteins in Multiple Sclerosis Pathogenesis: Friend or Foe?

Multiple Sclerosis is a complex chronic inflammatory, neurodegenerative disease conditioned by genetic, epigenetic and environmental factors. Main pathological features of MS include areas of focal demyelination of white matter characterized by gliosis, neuron and oligodendrocyte loss. Neurodegenerative as well as immune-mediated processes play a role in the pathogenesis of this disease. One of these immunogenic factors could be represented by the heat shock proteins. HSP exhibit cytoprotective and cytostimulatory effects due to their molecular chaperones role, in many brain model misfolding diseases such as Alzheimer’s and Parkinson’s diseases, whereas still no unambiguous results have bee…

La regolazione dell’espressione della MMP2 in mesoangioblasti è dipendente dai livelli della proteina Hsp70.

Hsp70 release from mesoangioblast A6 stem cells through vesicles

Hsp70 localizes differently from chaperone Hsc70 in mouse mesoangioblasts under physiological growth conditions

Mouse A6 mesoangioblasts express Hsp70 even in the absence of cellular stress. Its expression and its intracellular localization were investigated under normal growth conditions and under hyperthermic stress. Immunofluorescence assays indicated that without any stress a fraction of Hsp70 co-localized with actin microfilaments, in the cell cortex and in the contractile ring of dividing cells, while the Hsc70 chaperone did not. Hsp70 immunoprecipitation assays confirmed that a portion of Hsp70 binds actin. Immunoblot assays showed that both proteins were present in the nucleus. After heat treatment Hsp70 and actin continued to co-localize in the leading edge of A6 cells but not on microfilame…

Hsp70 function inside and outside mouse mesoangioblast stem cells

Isolation of cell clones from stem cell population more resistant to oxidative stress for tissue repair.

A6 stem cells release vesicles containing HSP70i

Role of Hsp70 in Multiple Sclerosis: An Overview

For many years heat shock protein 70 (Hsp70) was considered exclusively an intracellular chaperone contributing to protein proteostasis and in apoptotic pathway block. Lately it has been demonstrated that Hsp70 is actively released in the extracellular environment, thereby promoting the activation of the immune system by stimulating innate and adaptive responses through the activation of APCs. Its expression in the nervous system is induced in a variety of pathological conditions. Emerging evidences displayed that Hsp70 is a critical regulator in normal neural cells. Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS) directed against myelin antigens. In thi…

Armazon celulary organelos: citoesqueleto, mitocondria, peroxisomas y nucleo

MMP2 expression in mouse mesoangioblast is dependent on Hsp70 level.

Sperm activation and sperm-egg interaction

Different steps of sperm activation such as acrosomal reaction and capacitation are described in details. The molecules involved in sperm-egg interaction are also reported.

Hsp70 is required for optimal cell proliferation in mouse A6 mesoangioblast stem cells.

Mouse Hsp70 (70 kDa heat shock protein) is preferentially induced by heat or stress stimuli. We previously found that Hsp70 is constitutively expressed in A6 mouse mesoangioblast stem cells, but its possible role in these cells and the control of its basal transcription remained unexplored. Here we report that in the absence of stress, Ku factor is able to bind the HSE (heat shock element) consensus sequence in vitro, and in vivo it is bound to the proximal hsp70 promoter. In addition, we show that constitutive hsp70 transcription depends on the co-operative interaction of different factors such as Sp1 (specificity protein 1) and GAGA-binding protein with Ku factor, which binds the HSE cons…

Mouse mesoangioblasts release Hsp70 in a controlled manner through membrane vesicle shedding

Mesoangioblaststs (Mabs) are mesodermal stem cells associated with vessels which can differentiate into different mesoderm cell types. They have the property to cross endothelial barrier and when injected into circulation they localize into damaged tissues. A6 cells, a clone of mouse Mabs, express Hsp70 protein in physiological conditions and this synthesis may be required to answer to several intra- and/or extra-cellular needs. It was found that Hsp70 may be released outside from several type of cells, but its role remains still undefined. In order to clarify the reason of Hsp70 constitutive expression, we prepared several A6 clones with different levels of Hsp70 expression by stable RNA i…

Role and regulation of HSP70 in A6 mesoangioblast stem cells

Vanadium Modulates Proteolytic Activities and MMP-14-Like Levels during Paracentrotus lividus Embryogenesis

The increasing industrial use of vanadium (V), as well as its recent medical use in various pathologies has intensified its environmental release, making it an emerging pollutant. The sea urchin embryo has long been used to study the effects induced by metals, including V. In this study we used an integrated approach that correlates the biological effects on embryo development with proteolytic activities of gelatinases that could better reflect any metal-induced imbalances. V-exposure caused morphological/morphometric aberrations, mainly concerning the correct distribution of embryonic cells, the development of the skeleton, and the embryo volume. Moreover, V induced a concentration change …

Differences in intercellular communication during clinical relapse and gadolinium-enhanced MRI in patients with relapsing remitting multiple sclerosis: A study of the composition of extracellular vesicles in cerebrospinal fluid

This study was designed based on the hypothesis that changes in both the levels and surface marker expression of extracellular vesicles (EVs) isolated from the cerebrospinal fluid (CSF) may be associated with the clinical form, disease activity, and severity of multiple sclerosis (MS). The analyzes were performed on subjects affected by MS or other neurological disorders. EVs, which were isolated by ultracentrifugation of CSF samples, were characterized by flow cytometry. A panel of fluorescent antibodies was used to identify the EV origin: CD4, CCR3, CCR5, CD19, and CD200, as well as isolectin IB4. The Mann–Whitney U-test and Kruskal–Wallis test were used for statistical analyzes. EVs isol…

Double Face of eHsp70 in Front of Different Situations. Multiple Role of eHsp70

The Hsp70 family is one of the best conserved and abundant member of the heat shock proteins (HSP). This family includes several members and in particular one constitutively expressed member (Hsc70) and another one inducibly expressed under several stress conditions (Hsp70). To date, the intracellular functions of Hsp70 are well defined, and increasing evidences establish its roles in the extracellular environment, such as cytoprotection and immunomodulation. Increasing evidences suggest that several cell types are able to release Hsp70 in the extracellular environment, both under physiological and stress conditions. At the same time many release mechanisms have been identified. This chapte…

A sub-population of mesoangioblasts displays features of resistance and proliferation confirmed by transcriptome analysis.

A6 stem cells culture into a biodegradable PLLA scaffold

Factors which influence sperm ability to fertilize.

Different facrors influence animal sperm abiÌiry ro fertilizc- Some ofthem are revieu'ed here, spermmotiliry block to polyspermy, chemioattracrìon, sperm cornpetition lor fcrtilization. Old and new data are report-ed, as lor exanplc the new notions on sperm moriliry derived from sitc directed mutagenesis in rodents, the newnotions on thc odour receptors in m:rrnmalian sperm attr-action and ncw notions on sperm competìtion, which isvariable in diflerent species

The migration capability of mouse mesoangioblast stem cells depends on Hsp autocrine signalling.

Extracellular vesicles isolated by cerebrospinal fluid as biomarkers of inflammation in multiple sclerosis and inflammatory neurological diseases.

Paracrine effect of membrane vesicles released by mouse mesoangioblast stem cells

Introduction: Mouse mesoangioblasts are vessel-associated multipotent progenitor stem cells, which are able to differentiate into different mesodermal cell types. In our previous paper, we have demonstrated that mesoangioblasts are able to shed in the extracellular environment membrane vesicles (EVs), which contain both structural proteins and biological factors such as FGF2 and the two gelatinases MMP2/9. We investigated whether these EV interact in a paracrine way with other cell types different from mesoangioblasts, and eventually the effects of this interaction. Methods: Mesoangioblast EVs were collected from conditioned media by ultracentrifugation. Total mRNAs from mesoangioblasts and…

Mesoangioblast MMP2 regulation is dependent on Hsp70 level

Protective role of heat shock proteins in Parkinson's disease.

Parkinson’s disease (PD) is the second most common neurodegenerative disease after Alzheimer’s disease. Despite a large amount of research, the pathogenetic mechanism of these diseases has not yet been clarified. Abnormal protein folding, oxidative stress, mitochondrial dysfunction, and apoptotic mechanisms have all been reported as causes of neurodegenerative diseases in association with neuroinflammatory mechanisms which, by generating deleterious molecules, could promote the cascade of events leading to neurodegeneration. Heat shock proteins (HSPs) play a central role in preventing protein misfolding and inhibiting apoptotic activity, and represent a class of proteins potentially involve…

Stress response in mesoangioblast stem cells

Stem cells are presumed to survive various stresses, since they are recruited to areas of tissue damage and regeneration, where inflammatory cytokines and cytotoxic cells may result in severe cell injury. We explored the ability of mesoangioblasts to respond to different cell stresses such as heat, heavy metals and osmotic stress, by analyzing heat shock protein (HSP)70 synthesis as a stress indicator. We found that the A6 mesoangioblast stem cells constitutively synthesize HSP70 in a heat shock transcription factor (HSF)-independent way. However, A6 respond to heat shock and cadmium treatment by synthesizing HSP70 over the constitutive expression and this synthesis is HSF1 dependent. The e…

Synthesis of a porous and biodegradable PLLA scaffold for application of tissue engineering

Analisi dell’espressione dell’HSP70 inducubile in cellule staminali ed in tessutiu adulti di topo.

Effects of H2O2 on mesoangioblast stem cells: survival and cell death

EFFECT OF THE IMPase INHIBITOR L690,330 ON SEA URCHIN DEVELOPMENT

Abstract A variety of concentrations of the IMPase inhibitor L690,330 were added to sea urchin embryos. Immediate arrest of development was obtained for concentrations from 7.5 m m on. Concentrations lower than 3.5 m m permitted gastrulation but inhibited skeletogenesis and disturbed elongation along the animal–vegetal axis. The latter results are similar to those obtained by counteracting lithium effect with myoinositol, which are suggested to be due to partial relief of IMPase inhibition.

WIN55,212-2-induced expression of Mir-29b1 favours the suppression of osteosarcoma cell migration in a SPARC-independent manner

WIN55,212-2 (WIN) is a synthetic agonist of cannabinoid receptors that displays promising antitumour properties. The aim of this study is to demonstrate that WIN is able to block the migratory ability of osteosarcoma cells and characterize the mechanisms involved. Using wound healing assay and zymography, we showed that WIN affects cell migration and reduces the activity of the metalloproteases MMP2 and MMP9. This effect seemed to be independent of secreted protein acidic and rich in cysteine (SPARC), a matricellular protein involved in tissue remodeling and extracellular matrix deposition. SPARC release was indeed prevented by WIN, and SPARC silencing by RNA interference did not influence …

WIN-induced vesiculation cooperates to the inhibition of osteosarcoma cell migration

Introduction. Animal cells release vesicles that mediate the secretion of a variety of factors in the surrounding environment affecting neighboring cells. There is increasing evidence that secreted vesicles play an important role as vehicle of intercellular communication in different biological systems and are able to influence both physiological and pathological processes. Recently, we have reported that the synthetic cannabinoid WIN55,512 is able to induce osteosarcoma MG63 cell death and negatively affect cell migration. Here, we study the effects of WIN on the induction of vesicle secretion and their possible role in WIN-dependent reduction of osteosarcoma cell migratory ability. Method…

Shedding of vesicles from mesoangioblast A6 stem cells

Mesoangioblast behaviour in oxidative stress condition

Zymographic Analysis of Matrix Metalloproteinases (MMP-2 and MMP-9) in Cerebrospinal Fluid and Sera from Patients with Multiple Sclerosis

Hsp70 in mesoangioblast A6 stem cells.

Mesenchymal and Induced Pluripotent Stem Cells-Derived Extracellular Vesicles: The New Frontier for Regenerative Medicine?

Regenerative medicine aims to repair damaged, tissues or organs for the treatment of various diseases, which have been poorly managed with conventional drugs and medical procedures. To date, multimodal regenerative methods include transplant of healthy organs, tissues, or cells, body stimulation to activate a self-healing response in damaged tissues, as well as the combined use of cells and bio-degradable scaffold to obtain functional tissues. Certainly, stem cells are promising tools in regenerative medicine due to their ability to induce de novo tissue formation and/or promote organ repair and regeneration. Currently, several studies have shown that the beneficial stem cell effects, espec…

Extracellular Hsp70 Enhances Mesoangioblast Migration via an Autocrine Signaling Pathway

Mouse mesoangioblasts are vessel-associated progenitor stem cells endowed with the ability of multipotent mesoderm differentiation. Therefore, they represent a promising tool in the regeneration of injured tissues. Several studies have demonstrated that homing of mesoangioblasts into blood and injured tissues are mainly controlled by cytokines/chemokines and other inflammatory factors. However, little is known about the molecular mechanisms regulating their ability to traverse the extracellular matrix (ECM). Here, we demonstrate that membrane vesicles released by mesoangioblasts contain Hsp70, and that the released Hsp70 is able to interact by an autocrine mechanism with Toll-like receptor …

Extracellular Hsp70 Enhances Mesoangioblast Migration via an Autocrine Signaling Pathway

Mouse mesoangioblasts are vessel-associated progenitor stem cells endowed with the ability of multipotent mesoderm differentiation. Therefore, they represent a promising tool in the regeneration of injured tissues. Several studies have demonstrated that homing of mesoangioblasts into blood and injured tissues are mainly controlled by cytokines/chemokines and other inflammatory factors. However, little is known about the molecular mechanisms regulating their ability to traverse the extracellular matrix (ECM). Here, we demonstrate that membrane vesicles released by mesoangioblasts contain Hsp70, and that the released Hsp70 is able to interact by an autocrine mechanism with Toll-like receptor …

Porous PLLA scaffolds are optimal substrates for internal colonization by A6 mesoangioblasts and immunocytochemical analyses

In the present paper, mouse mesoangioblasts were seeded onto bidimensional matrices and within three-dimensional porous scaffolds of poly(L-lactic acid) (PLLA), in the presence or absence of type I collagen coating, observed under the scanning electron microscope, and tested for their adhesion, survival and proliferation. Immunolocalization of Hsp70, an abundant and ubiquitous intracellular protein in these cells, was also performed in sectioned cell-containing scaffolds under the confocal fluorescence microscope to check whether "in situ" analysis of intracellular constituents was feasible. The data obtained show that PLLA films allow direct cell adhesion and represent an optimal support f…

Ciclo celular, apoptosis y envejecimiento

H2O2 resistant mesoangioblast clone isolation with a distinct survival advantage in vitro and in vivo

The release of molecules from damaged tissues stimulates both resident and circulating stem cells to initiate a tissue repair programme. 1 However, during transplantation procedures the therapeutic efficacy of stem cells is compromised by reduced homing capability towards the target site.2,3 Furthermore, cell survival is very low and many studies focused on improving cell viability upon implantation. In this study, we performed in vitrosevere oxidative stress to select some more resistant mouse mesoangioblasts, vessel-associated progenitor stem cells endowed with the ability of multipotent mesoderm differentiation. We found that the selected subpopulation retains selfrenewal and myogenic di…

Autophagy and apoptosis regolate survival of mesoangioblast stem cells subjected to oxidative stress

Extracellular vesicles in multiple sclerosis as possible biomarkers: Dream or reality?

Extracellular vesicles are recently described as specialized structures for intercellular communication. Their role in the central nervous system was diffusely studied in both physiological and pathological condition. In particular, an increased extracellular vesicle number was detected in several autoimmune diseases, including multiple sclerosis, a chronic autoimmune, inflammatory, demyelinating and neurodegenerative disease. This chapter summarizes the available information on the involvement of the extracellular vesicles in multiple sclerosis pathogenesis and their possible use as biomarker of therapy efficacy.

Mesoangioblast A6 stem cells release vesicles containing the inducible HSP70

An hsf2-like factor is present in the invertebrates: Characterization and purification in sea urchin embryos and its localization in primary mesenchime cells

Ku factor is responsible of Hsp70 basal transcription in mouse mesoangioblasts

A homemade cytospin apparatus

Inducible HSP70 localization in mesoangioblast A6 stem cells

Hsp70 is involved in MMP2 level in mouse mesoangioblast stem cells

Double Face of eHsp70 in Front of Different Situations

The Hsp70 family is one of the best conserved and abundant member of the heat shock proteins (HSP). This family includes several members and in particular one constitutively expressed member (Hsc70) and another one inducibly expressed under several stress conditions (Hsp70). To date, the intracellular functions of Hsp70 are well defined, and increasing evidences establish its roles in the extracellular environment, such as cytoprotection and immunomodulation. Increasing evidences suggest that several cell types are able to release Hsp70 in the extracellular environment, both under physiological and stress conditions. At the same time many release mechanisms have been identified. This chapte…

Extracellular membrane vesicles as a mechanism of cell-to-cell communication: advantages and disadvantages.

Microvesicles represent a newly identified mechanism of intercellular communication. Two different types of microvesicles have been identified: membrane-derived vesicles (EVs) and exosomes. EVs originate by direct budding from the plasma membrane, while exosomes arise from ectocytosis of multivesicular bodies. Recent attention has focused on the capacity of EVs to alter the phenotype of neighboring cells to make them resemble EV-producing cells. Stem cells are an abundant source of EVs, and the interaction between stem cells and the microenvironment (i.e., stem cell niche) plays a critical role in determining stem cell phenotype. The stem cell niche hypothesis predicts that stem cell number…

Cell clones more resistant to oxidative stress selecte from embryonic and adult mesoangioblast stem cell population.

Transcriptome analysis after a strong oxidative stress highlighted a mesoangioblast stem cell sub-population with important different capability

Rapid changes in heat-shock cognate 70 levels, heat-shock cognate phosphorylation state, heat-shock transcription factor, and metal transcription factor activity levels in response to heavy metal exposure during sea urchin embryonic development.

The aim of the present study was to analyze and compare the effects of several metals on the embryos of the sea urchin Paracentrotus lividus, a key species within the Mediterranean Sea ecosystem. Embryos were continuously exposed from fertilization to the following metals: 0.6 mg/l copper, 3 mg/l lead, and 6 mg/l nickel. The embryos were then monitored for metal responses at the gastrula stage, which occurred 24 h after exposure. A biochemical multi-experimental approach was taken and involved the investigation of the levels of HSC70 expression and the involvement of heat shock factor (HSF) and/or metal transcription factor (MTF) in the response. Immunoblotting assays and electrophoretic mo…

Proteolytic activity and MMP-14-like protein levels are affected by Vanadium in Paracentrotus lividus Embryo

The increasing industrial use of vanadium (V), as well as its recent medical use in various pathologies has intensified its environmental release, making it an emerging pollutant [1]. The sea urchin embryo has long been used to study the effects induced by metals, including V. In this study we used an integrated approach that correlates the biological effects on embryo development with proteolytic activities of gelatinases that could better reflect any metal induced imbalances. V-exposure caused morphological/morphometric aberrations, mainly concerning the correct dis-tribution of embryonic cells, the development of the skeleton and the embryo volume [2]. Moreover, V induced a concentration…

Regeneration in invertebrates

The mechanisms of regeneration are reviewed from a genetic, cytological and molecular biological points of view. Planarians and Hydra have been chosen and illustrated as biological examples.