Advances in Regenerative medicine
Book published by Intech-open access publisher (2011)
PREFACE In order to better introduce this book, it is important to define regenerative medicine as this field is built through a combination of multiple elements including living cells, matrix to support ... [more ▼]
PREFACE In order to better introduce this book, it is important to define regenerative medicine as this field is built through a combination of multiple elements including living cells, matrix to support the living cells (i.e. a scaffold), and cell communicators (or signaling systems) to stimulate the cells, and their surrounding environment to grow and develop into new tissue or organ. Indeed, regenerative medicine is an emerging multidisciplinary field involving biology, medicine, and engineering that is likely to revolutionize the ways we improve the health and quality of life for millions of people worldwide by restoring, maintaining, or enhancing tissue and organ function. Even if the origins of regenerative medicine can be found in Greek mythology, as attested by the story of Prometheus, the Greek god whose immortal liver was feasted on day after day by Zeus' eagle; many challenges persist in order to successfully regenerate lost cells, tissues or organs and rebuilt all connections and functions. In this book, we will cover few aspects of regenerative medicine highlighting major advances and remaining challenges in cellular therapy (including cell communicators) and tissue/organ engineering. CELL REPLACEMENT THERAPY The types of cells that are used are dependent on the type of tissue that needs to be repaired. Several cells have been suggested as suitable for cellular therapies: i.e. embryonic stem cells (ES), induced pluripotent stem cells (iPS); somatic stem cells from fetal or adult tissues. The potential use of fetal tissue or differentiated embryonic stem cells from allogenic sources suffer limitations due to tissue availability, ethical issues or safety concerns. On the contrary, adult somatic stem cells can be used in autologus graft procedure, avoiding patient’s immunosuppression. In this book, several chapters will be developed on stem cell applications in regenerative medicine focusing on several organs or tissues like brain, heart, liver or retina. CELL COMMUNICATORS. The circulatory system is involved in the transport of a wide variety of biological molecules and cells and can be considered as the body's basic communication system. Cell communicators act as a signaling system, which stimulates the cells into action. In some cases those communicators could lead cells to integrate damage tissues and rebuilt lost connections, however, some signals could also induce cellular stress responses conducting to cell death. Few of those aspects will be directly address in this book. TISSUE ENGINEERING OR …WHERE BIOLOGY MEETS ENGINEERING. All cells within tissues are separated and interlinked by a matrix or structure. The consistency of the matrix may vary from a liquid, as in blood; to semi-solid, as in cartilage; to a solid, as in bone. Tissue engineers either implant cells into a matrix or create the proper conditions for the living cells to build their own three dimensional matrix. Such a matrix provides the structure that supports the cells and creates the physiological environment for them to interact within the host tissue. The success or failure of an implant material in the body depends on a complex interaction between a synthetic ‘foreign body’ and the ‘host tissue’, which involve not only biological, but also mechanical, physical and chemical mediated factors. The latest advance in tissue engineering will be developed in this book underlying many challenges that remain pending in this field. [less ▲]Detailed reference viewed: 174 (10 ULg)
Adult bone marrow: which stem cells for cellular therapy protocols in neurodegenerative disorders?
Wislet-Gendebien, Sabine ; Laudet, Emerence ; Neirinckx, Virginie et al
in Journal of Biomedicine & Biotechnology (2011)
The generation of neuronal cells from stem cells obtained from adult bone marrow is of significant clinical interest in order to design new cell therapy protocols for several neurological disorders. The ... [more ▼]
The generation of neuronal cells from stem cells obtained from adult bone marrow is of significant clinical interest in order to design new cell therapy protocols for several neurological disorders. The recent identification in adult bone marrow of stem cells derived from the neural crests (NCSC) might explain the neuronal phenotypic plasticity shown by bone marrow cells. However, little information is available about the nature of these cells compared to mesenchymal stem cells (MSC). In this manuscript, we will review all information available concerning NCSC from adult tissues and their possible use in regenerative medecine. Moreover, as multiple recent studies showed the beneficial effect of bone marrow stromal cells in neurodegenerative diseases, we will discuss which stem cells isolated from adult bone marrow should be more suitable for cell replacement therapy. [less ▲]Detailed reference viewed: 49 (11 ULg)
Effect of Ser-129 Phosphorylation on Interaction of Alpha-Synuclein with Synaptic and Cellular Membranes.
Wislet-Gendebien, Sabine ; ; et al
in Journal of Biological chemistry (2011), 286(41), 35863-73
In the healthy brain, less than 5% of α-synuclein (α-syn) is phosphorylated at serine 129 (Ser(P)-129). However, within Parkinson disease (PD) Lewy bodies, 89% of α-syn is Ser(P)-129. The effects of Ser(P ... [more ▼]
In the healthy brain, less than 5% of α-synuclein (α-syn) is phosphorylated at serine 129 (Ser(P)-129). However, within Parkinson disease (PD) Lewy bodies, 89% of α-syn is Ser(P)-129. The effects of Ser(P)-129 modification on α-syn distribution and solubility are poorly understood. As α-syn normally exists in both membrane-bound and cytosolic compartments, we examined the binding and dissociation of Ser(P)-129 α-syn and analyzed the effects of manipulating Ser(P)-129 levels on α-syn membrane interactions using synaptosomal membranes and neural precursor cells from α-syn-deficient mice or transgenic mice expressing human α-syn. We first evaluated the recovery of the Ser(P)-129 epitope following either α-syn membrane binding or dissociation. We demonstrate a rapid turnover of Ser(P)-129 during both binding to and dissociation from synaptic membranes. Although the membrane binding of WT α-syn was insensitive to modulation of Ser(P)-129 levels by multiple strategies (the use of phosphomimic S129D and nonphosphorylated S129A α-syn mutants; by enzymatic dephosphorylation of Ser(P)-129 or proteasome inhibitor-induced elevation in Ser(P)-129; or by inhibition or stable overexpression of PLK2), PD mutant Ser(P)-129 α-syn showed a preferential membrane association compared with WT Ser(P)-129 α-syn. Collectively, these data suggest that phosphorylation at Ser-129 is dynamic and that the subcellular distribution of α-syn bearing PD-linked mutations, A30P or A53T, is influenced by the phosphorylation state of Ser-129. [less ▲]Detailed reference viewed: 40 (7 ULg)
Wnt1 and BMP2: two factors recruiting multipotent neural crest progenitors isolated from adult bone marrow
Glejzer, Aneta ; Laudet, Emerence ; Leprince, Pierre et al
in Cellular and Molecular Life Sciences : CMLS (2011), 68/12
Recent studies have shown that neural crestderived progenitor cells can be found in diverse mammalian tissues including tissues that were not previously shown to contain neural crest derivatives, such as ... [more ▼]
Recent studies have shown that neural crestderived progenitor cells can be found in diverse mammalian tissues including tissues that were not previously shown to contain neural crest derivatives, such as bone marrow. The identification of those ‘‘new’’ neural crest-derived progenitor cells opens new strategies for developing autologous cell replacement therapies in regenerative medicine. However, their potential use is still a challenge as only few neural crest-derived progenitor cells were found in those new accessible locations. In this study, we developed a protocol, based on wnt1 and BMP2 effects, to enrich neural crest-derived cells from adult bone marrow. Those two factors are known to maintain and stimulate the proliferation of embryonic neural crest stem cells, however, their effects have never been characterized on neural crest cells isolated from adult tissues. Using multiple strategies from microarray to 2D-DIGE proteomic analyses, we characterized those recruited neural crest-derived cells, defining their identity and their differentiating abilities. [less ▲]Detailed reference viewed: 109 (35 ULg)
Differential regulation of wild-type and mutant alpha-synuclein binding to synaptic membranes by cytosolic factors.
Wislet-Gendebien, Sabine ; ; et al
in BMC Neuroscience (2008), 9
BACKGROUND: Alpha-Synuclein (alpha-syn), a 140 amino acid protein associated with presynaptic membranes in brain, is a major constituent of Lewy bodies in Parkinson's disease (PD). Three missense ... [more ▼]
BACKGROUND: Alpha-Synuclein (alpha-syn), a 140 amino acid protein associated with presynaptic membranes in brain, is a major constituent of Lewy bodies in Parkinson's disease (PD). Three missense mutations (A30P, A53T and E46K) in the alpha-syn gene are associated with rare autosomal dominant forms of familial PD. However, the regulation of alpha-syn's cellular localization in neurons and the effects of the PD-linked mutations are poorly understood. RESULTS: In the present study, we analysed the ability of cytosolic factors to regulate alpha-syn binding to synaptic membranes. We show that co-incubation with brain cytosol significantly increases the membrane binding of normal and PD-linked mutant alpha-syn. To characterize cytosolic factor(s) that modulate alpha-syn binding properties, we investigated the ability of proteins, lipids, ATP and calcium to modulate alpha-syn membrane interactions. We report that lipids and ATP are two of the principal cytosolic components that modulate Wt and A53T alpha-syn binding to the synaptic membrane. We further show that 1-O-hexadecyl-2-acetyl-sn-glycero-3-phosphocholine (C16:0 PAF) is one of the principal lipids found in complex with cytosolic proteins and is required to enhance alpha-syn interaction with synaptic membrane. In addition, the impaired membrane binding observed for A30P alpha-syn was significantly mitigated by the presence of protease-sensitive factors in brain cytosol. CONCLUSION: These findings suggest that endogenous brain cytosolic factors regulate Wt and mutant alpha-syn membrane binding, and could represent potential targets to influence alpha-syn solubility in brain. [less ▲]Detailed reference viewed: 41 (8 ULg)
Stem cell factor and mesenchymal and neural stem cell transplantation in a rat model of Huntington's disease.
; ; et al
in Molecular & Cellular Neuroscience [=MCN] (2008), 37(3), 454-70
Neural and mesenchymal stem cells have been proposed as alternative sources of cells for transplantation into the brain in neurodegenerative disorders. However, the endogenous factors controlling their ... [more ▼]
Neural and mesenchymal stem cells have been proposed as alternative sources of cells for transplantation into the brain in neurodegenerative disorders. However, the endogenous factors controlling their engraftment within the injured parenchyma remain ill-defined. Here, we demonstrate significant engraftment of undifferentiated exogenous mesenchymal or neural stem cells throughout the lesioned area in a rat model for Huntington's disease, as late as 8 weeks post-transplantation. We show that stem cell factor (SCF), strongly up-regulated within host cells in the damaged striatum, is able to activate the SCF receptor c-kit and its signaling pathway and to promote the migration and proliferation of mesenchymal and neural stem cells in vitro. Furthermore, c-kit receptor blockade alters neural stem cell distribution within the lesioned striatum. Host SCF expression is observed in atypical cells expressing glial fibrillary acidic protein and doublecortin in the lesioned striatum and in migrating doublecortin-positive progenitors. Taken together, these data demonstrate that SCF produced in situ in the lesioned striatum is an important factor in promoting the engraftment of stem cells within the lesioned brain. [less ▲]Detailed reference viewed: 121 (7 ULg)
Regulation of nestin expression by thrombin and cell density in cultures of bone mesenchymal stem cells and radial glial cells.
Wislet-Gendebien, Sabine ; ; et al
in BMC Neuroscience (2007), 8
BACKGROUND: Bone marrow stromal cells and radial glia are two stem cell types with neural phenotypic plasticity. Bone marrow mesenchymal stem cells can differentiate into osteocytes, chondrocytes and ... [more ▼]
BACKGROUND: Bone marrow stromal cells and radial glia are two stem cell types with neural phenotypic plasticity. Bone marrow mesenchymal stem cells can differentiate into osteocytes, chondrocytes and adipocytes, but can also differentiate into non-mesenchymal cell, i.e. neural cells in appropriate in vivo and in vitro experimental conditions. Likewise, radial glial cells are the progenitors of many neurons in the developing cortex, but can also generate astrocytes. Both cell types express nestin, an intermediate filament protein which is the hallmark of neural precursors. RESULTS: In this study, we demonstrate that thrombin, a multifunctional serine protease, stimulates the growth of radial glial cells (RG) and mesenchymal stem cells (MSCs) in a dose-dependent manner. In RG, the mitogenic effect of thrombin is correlated with increased expression of nestin but in MSCs, this mitogenic effect is associated with nestin down-regulation. Both cell types express the PAR-1 type receptor for Thrombin and the effect of Thrombin on both cell types can be mimicked by its analogue TRAP-6 activating specifically this receptor subtype or by serum which contains various amount of thrombin. Moreover, we also demonstrate that serum deprivation-induced expression of nestin in MSCs is inhibited by high cell density (> 50,000 cells/cm2). CONCLUSION: This work shows that thrombin stimulates the growth of both RG and MSCs and that nestin expression by MSCs and RG is regulated in opposite manner by thrombin in vitro. Thrombin effect is thus associated in both cell types with a proliferating, undifferentiated state but in RG this involves the induction of nestin expression, a marker of immaturity for neural progenitors. In MSCs however, nestin expression, as it corresponds to a progression from the mesenchymal "undifferentiated", proliferating phenotype toward acquisition of a neural fate, is inhibited by the mitogenic signal. [less ▲]Detailed reference viewed: 58 (8 ULg)
Cytosolic proteins regulate alpha-synuclein dissociation from presynaptic membranes.
Wislet-Gendebien, Sabine ; ; et al
in Journal of Biological Chemistry (2006), 281(43), 32148-55
Intracellular accumulation of insoluble alpha-synuclein in Lewy bodies is a key neuropathological trait of Parkinson disease (PD). Neither the normal function of alpha-synuclein nor the biochemical ... [more ▼]
Intracellular accumulation of insoluble alpha-synuclein in Lewy bodies is a key neuropathological trait of Parkinson disease (PD). Neither the normal function of alpha-synuclein nor the biochemical mechanisms that cause its deposition are understood, although both are likely influenced by the interaction of alpha-synuclein with vesicular membranes, either for a physiological role in vesicular trafficking or as a pathological seeding mechanism that exacerbates the propensity of alpha-synuclein to self-assemble into fibrils. In addition to the alpha-helical form that is peripherally-attached to vesicles, a substantial portion of alpha-synuclein is freely diffusible in the cytoplasm. The mechanisms controlling alpha-synuclein exchange between these compartments are unknown and the possibility that chronic dysregulation of membrane-bound and soluble alpha-synuclein pools may contribute to Lewy body pathology led us to search for cellular factors that can regulate alpha-synuclein membrane interactions. Here we reveal that dissociation of membrane-bound alpha-synuclein is dependent on brain-specific cytosolic proteins and insensitive to calcium or metabolic energy. Two PD-linked mutations (A30P and A53T) significantly increase the cytosol-dependent alpha-synuclein off-rate but have no effect on cytosol-independent dissociation. These results reveal a novel mechanism by which cytosolic brain proteins modulate alpha-synuclein interactions with intracellular membranes. Importantly, our finding that alpha-synuclein dissociation is up-regulated by both familial PD mutations implicates cytosolic cofactors in disease pathogenesis and as molecular targets to influence alpha-synuclein aggregation. [less ▲]Detailed reference viewed: 65 (4 ULg)
beta-carbolines induce apoptosis in cultured cerebellar granule neurons via the mitochondrial pathway
Hans, Grégory ; Malgrange, Brigitte ; et al
in Neuropharmacology (2005), 48(1), 105-117
N-Butyl-beta-carboline-3-carboxylate (betaCCB) is, together with 2-methyl-norharmanium and 2,9-dimethylnorharmanium ions, an endogenously occurring beta-carboline. Due to their structural similarities ... [more ▼]
N-Butyl-beta-carboline-3-carboxylate (betaCCB) is, together with 2-methyl-norharmanium and 2,9-dimethylnorharmanium ions, an endogenously occurring beta-carboline. Due to their structural similarities with the synthetic neurotoxin 1-methy14-phenyl-1,2,3,6-tetrahydropyridine (MPTP), harman and norharman compounds have been proposed to be involved in the pathogenesis of Parkinson's disease. While also structurally related, betaCCB has received much less interest in that respect although we had previously demonstrated that it induces the apoptotic cell death of cultured cerebellar granule neurons (CGNs). Herein, we have investigated the molecular events leading to CGN apoptosis upon betaCCB treatment. We first demonstrated that betaCCB-induced apoptosis occurs in neurons only, most likely as a consequence of a specific neuronal uptake as shown using binding/uptake experiments. Then we observed that, in betaCCB-treated CGNs, caspases 9, 3 and 8 were successively activated, suegesing an activation of the mitochondrial pathway. Consistently, betaCCB also induced the release from the mitochondrial intermembrane space of two pro-apoptotic factors. i.e. cytochrome c and apotptosis inducing factor (AIF). Interestingly, no mitochondrial membrane depolarisation was associated with this release. suggesting a mitochondrial permeability transition pore-independent mechanism. The absence of any neuroprotective effect provided by two mPTP inhibitors. i.e. cyclosporine A and bongkrekic acid. further supported this hypothesis. Together. these results show that betaCCB is specifically taken up by neuronal cells where it triggers a specific permeabilization of the outer mitochondrial membrane and a subsequent apoptotic cell death. (C) 2004 Elsevier Ltd. All rights reserved. [less ▲]Detailed reference viewed: 75 (14 ULg)
Astrocytic and neuronal fate of mesenchymal stem cells expressing nestin.
Wislet-Gendebien, Sabine ; Wautier, Franz ; Leprince, Pierre et al
in Brain Research Bulletin (2005), 68(1-2), 95-102
Classically, bone marrow mesenchymal stem cells (MSC) differentiate in vivo or in vitro into osteocytes, chondrocytes, fibroblasts and adipocytes. Recently, it was reported by several groups that MSC can ... [more ▼]
Classically, bone marrow mesenchymal stem cells (MSC) differentiate in vivo or in vitro into osteocytes, chondrocytes, fibroblasts and adipocytes. Recently, it was reported by several groups that MSC can also adopt a neural fate in appropriate in vivo or in vitro experimental conditions. However, it is unclear if those cells are really able to differentiate into functional neural cells and in particular into functional neurons. Some observations suggest that a cell fusion process underlies the neural fate adoption by MSC in vivo and first attempts to reproduce in vitro this neural fate decision in MSC cultures were unsuccessful. More recently, however, in several laboratories including ours, differentiation of MSC cultivated from adult rat bone marrow into astrocytes and neuron-like cells was demonstrated. More precisely, we stressed the importance of the expression by MSC of nestin, an intermediate filament protein associated with immaturity in the nervous system, as a pre-requisite to adopting an astrocytic or a neuronal fate in a co-culture paradigm. Using this approach, we have also demonstrated that the MSC-derived neuron-like cells exhibit several electrophysiological key properties classically devoted to neurons, including firing of action potentials. In this review, we will discuss the neurogenic potential of MSC, the factor(s) required for such plasticity, the molecular mechanism(s) underlying this neural plasticity, the importance of the environment of MSC to adopt this neural fate and the therapeutic potential of these observations. [less ▲]Detailed reference viewed: 61 (11 ULg)
Plasticity of cultured mesenchymal stem cells: switch from nestin-positive to excitable neuron-like phenotype.
Wislet-Gendebien, Sabine ; Hans, Grégory ; Leprince, Pierre et al
in Stem Cells (2005), 23(3), 392-402
Bone marrow mesenchymal stem cells (MSCs) can differentiate into several types of mesenchymal cells, including osteocytes, chondrocytes, and adipocytes, but, under appropriate experimental conditions, can ... [more ▼]
Bone marrow mesenchymal stem cells (MSCs) can differentiate into several types of mesenchymal cells, including osteocytes, chondrocytes, and adipocytes, but, under appropriate experimental conditions, can also differentiate into nonmesenchymal cells--for instance, neural cells. These observations have raised interest in the possible use of MSCs in cell therapy strategies for various neurological disorders. In the study reported here, we addressed the question of in vitro differentiation of MSCs into functional neurons. First, we demonstrate that when they are co-cultured with cerebellar granule neurons, adult MSCs can express neuronal markers. Two factors are needed for the emergence of neuronal differentiation of the MSCs: the first one is nestin expression by MSCs (nestin is a marker for the responsive character of MSCs to extrinsic signals), and the second one is a direct cell-cell interaction between neural cells and MSCs that allows the integration of these extrinsic signals. Three different approaches suggest that neural phenotypes arise from MSCs by a differentiation rather than a cell fusion process, although this last phenomenon can also coexist. The expression of several genes--including sox, pax, notch, delta, frizzled, and erbB--was analyzed by quantitative reverse transcription polymerase chain reaction (RT-PCR) in order to further characterize the nestin-positive phenotype compared to the nestin-negative one. An overexpression of sox2, sox10, pax6, fzd, erbB2, and erbB4 is found in nestin-positive MSCs. Finally, electrophysiological analyses demonstrate that MSC-derived neuron-like cells can fire single-action potentials and respond to several neurotransmitters such as GABA, glycine, and glutamate. We conclude that nestin-positive MSCs can differentiate in vitro into excitable neuron-like cells. [less ▲]Detailed reference viewed: 64 (8 ULg)
Etude des mécanismes moléculaires et cellulaires à la base de la plasticité phénotypique nerveuse des cellules souches mésenchymateuses.
Doctoral thesis (2004)Detailed reference viewed: 21 (2 ULg)
Nestin-positive mesenchymal stem cells favour the astroglial lineage in neural progenitors and stem cells by releasing active BMP4.
Wislet-Gendebien, Sabine ; Bruyere, Françoise ; Hans, Grégory et al
in BMC Neuroscience (2004), 5
BACKGROUND: Spontaneous repair is limited after CNS injury or degeneration because neurogenesis and axonal regrowth rarely occur in the adult brain. As a result, cell transplantation has raised much ... [more ▼]
BACKGROUND: Spontaneous repair is limited after CNS injury or degeneration because neurogenesis and axonal regrowth rarely occur in the adult brain. As a result, cell transplantation has raised much interest as potential treatment for patients with CNS lesions. Several types of cells have been considered as candidates for such cell transplantation and replacement therapies. Foetal brain tissue has already been shown to have significant effects in patients with Parkinson's disease. Clinical use of the foetal brain tissue is, however, limited by ethical and technical problems as it requires high numbers of grafted foetal cells and immunosuppression. Alternatively, several reports suggested that mesenchymal stem cells, isolated from adult bone marrow, are multipotent cells and could be used in autograft approach for replacement therapies. RESULTS: In this study, we addressed the question of the possible influence of mesenchymal stem cells on neural stem cell fate. We have previously reported that adult rat mesenchymal stem cells are able to express nestin in defined culture conditions (in the absence of serum and after 25 cell population doublings) and we report here that nestin-positive (but not nestin-negative) mesenchymal stem cells are able to favour the astroglial lineage in neural progenitors and stem cells cultivated from embryonic striatum. The increase of the number of GFAP-positive cells is associated with a significant decrease of the number of Tuj1- and O4-positive cells. Using quantitative RT-PCR, we demonstrate that mesenchymal stem cells express LIF, CNTF, BMP2 and BMP4 mRNAs, four cytokines known to play a role in astroglial fate decision. In this model, BMP4 is responsible for the astroglial stimulation and oligodendroglial inhibition, as 1) this cytokine is present in a biologically-active form only in nestin-positive mesenchymal stem cells conditioned medium and 2) anti-BMP4 antibodies inhibit the nestin-positive mesenchymal stem cells conditioned medium inducing effect on astrogliogenesis. CONCLUSIONS: When thinking carefully about mesenchymal stem cells as candidates for cellular therapy in neurological diseases, their effects on resident neural cell fate have to be considered. [less ▲]Detailed reference viewed: 86 (3 ULg)
Regulation of neural markers nestin and GFAP expression by cultivated bone marrow stromal cells.
Wislet-Gendebien, Sabine ; Leprince, Pierre ; Moonen, Gustave et al
in Journal of Cell Science (2003), 116(Pt 16), 3295-302
Bone marrow stromal cells can differentiate into many types of mesenchymal cells, i.e. osteocyte, chondrocyte and adipocyte, but can also differentiate into non-mesenchymal cells, i.e. neural cells under ... [more ▼]
Bone marrow stromal cells can differentiate into many types of mesenchymal cells, i.e. osteocyte, chondrocyte and adipocyte, but can also differentiate into non-mesenchymal cells, i.e. neural cells under appropriate in vivo experimental conditions (Kopen et al., 1999; Brazelton et al., 2000; Mezey et al., 2000). This neural phenotypic plasticity allows us to consider the utilization of mesenchymal stem cells as cellular material in regenerative medicine. In this study, we demonstrate that cultured adult rat stromal cells can express nestin, an intermediate filament protein predominantly expressed by neural stem cells. Two factors contribute to the regulation of nestin expression by rat stromal cells: serum in the culture medium inhibits nestin expression and a threshold number of passages must be reached below which nestin expression does not occur. Only nestin-positive rat stromal cells are able to form spheres when they are placed in the culture conditions used for neural stem cells. Likewise, only nestin-positive stromal cells are able to differentiate into GFAP (glial fibrillary acidic protein)-positive cells when they are co-cultivated with neural stem cells. We thus demonstrated that adult rat stromal cells in culture express nestin in absence of serum after passaging the cells at least ten times, and we suggest that nestin expression by these cells might be a prerequisite for the acquisition of the capacity to progress towards the neural lineage. [less ▲]Detailed reference viewed: 104 (4 ULg)
Régulation de l’expression de la nestine par les cellules souches mésenchymateuses en culture
Master of advanced studies dissertation (2002)Detailed reference viewed: 53 (4 ULg)
Nestin expression in cultivated mesenchymal stem cells: Regulation and potential role in their neural differentiation
Wislet-Gendebien, Sabine ; Leprince, Pierre ; Moonen, Gustave et al
in Glia (2002, May), (Suppl. 1), 87
Bone marrow stromal cells can differentiate into many types of mesenchymal cells, i.e. osteocyte, chondrocyte, fibroblast and adipocyte, but can also differentiate into non-mesenchymal cell, i.e. neural ... [more ▼]
Bone marrow stromal cells can differentiate into many types of mesenchymal cells, i.e. osteocyte, chondrocyte, fibroblast and adipocyte, but can also differentiate into non-mesenchymal cell, i.e. neural cells in appropriate in vivo experimental conditions (Kopen and al.,PNAS,96, 10711,1999, Brazelton and al, Science, 290,1175, 2000, Mezey and al, Science, 290,1179, 2000). In neurological disorders, such as Alzheimer's and Parkinson's diseases, auto-transplantation of neural cell types derived from mesenchymal stem cells offers the potential of replacing lost cells and recovering lost functions. Nestin is an intermediate filament protein predominantly expressed by neural stem cells and is used to identify neural progenitor. In this study, we demonstrate that cultured rat mesenchymal stem cells (rMSC) can express nestin in appropriate conditions. Two factors contribute to the regulation of nestin expression by rMSC : 1) the presence of serum-derived components in the culture medium which repress nestin expression and 2) the cell’s number of passages. LPA and thrombin mimic this serum effect. Furthermore, when nestin- positive cells are trypsinized and resuspended into culture conditions used for neural stem cells (NSC), sphere formation is observed. Likewise, by co-cultivating nestin-positive rMSC with NSC derived from green mouse, heterogenous spheres were obtained. When those heterogenous spheres are placed on polyornithine-coated surfaces, a differentiation of some rMSC into GFAP-positive cells occurs. These results indicate that nestin expression might be a pre-requisite for the acquisition by rMSC of the capacity to differentiate into some neural cell types. [less ▲]Detailed reference viewed: 161 (4 ULg)
Clonage and expression de la protéine chimère TEM-RIM2
Master's dissertation (2000)Detailed reference viewed: 26 (2 ULg)