BRITISH JOURNAL OF HAEMATOLOGY, Issue 4 2010
Hady Felfly
Summary Minimal criteria requirements of stem cell replacement, conditioning regimen and modalities of infusion essential for cure of sickle cell disease (SCD) by bone marrow(BM)/stem cell transplantation or gene therapy must be established prior to clinical trials. The threshold of normal BM/stem cells for therapeutic correction of this red blood cell disorder was evaluated in the SAD murine SCD model from peripheral donor white blood cells. From 11 groups of stable chimeric SAD mice (5,92%) analyzed over ,2 years, mice with ,16% normal donor stem cells showed improvement of haematological and erythroid responses. Mice in the 26% chimeric group and above demonstrated substantial amelioration of organ pathologies with generalized decreased iron deposits, fibrosis and reached normal lifespan. Subsequently, the minimal myelosuppression concurrently with number and timing of infusions and number of BM cells was determined to reach therapeutic threshold in SAD mice. Higher myelosuppression (2 Gy vs. 1 Gy) and cell number in single infusion led to increased chimerism. Importantly, administration of three-equivalent cell subdoses within 28 h of mild myelosuppression resulted in 100% recipient engraftment at therapeutic levels. These studies established the long-term therapeutic chimeric threshold of normal white blood cells at ,26% and determined the minimal fractionated BM/stem cell doses concomitant with mild myelosuppression for significant correction of SCD in SAD mice. [source]

ADDRESS OF HIS HOLINESS POPE BENEDICT XVI TO PARTICIPANTS OF THE SYMPOSIUM ON THE THEME: ,STEM CELLS: WHAT FUTURE FOR THERAPY?

CELL PROLIFERATION, Issue 2008
SCIENTIFIC ASPECTS AND BIOETHICAL PROBLEMS'
No abstract is available for this article. [source]

Tissue Engineering: Stem Cell Aligned Growth Induced by CeO2 Nanoparticles in PLGA Scaffolds with Improved Bioactivity for Regenerative Medicine (Adv. Funct.

ADVANCED FUNCTIONAL MATERIALS, Issue 10 2010
Mater.
[source]

Stem Cell Aligned Growth Induced by CeO2 Nanoparticles in PLGA Scaffolds with Improved Bioactivity for Regenerative Medicine

ADVANCED FUNCTIONAL MATERIALS, Issue 10 2010
Corrado Mandoli
Abstract Hybrid 2D polymeric,ceramic biosupports are fabricated by mixing a nanostructured CeO2 powder with 85:15 poly(D,L -lactic- co -glycolic acid) (PLGA)/dichloromethane solutions at specific concentrations, followed by solvent casting onto pre-patterned molds. The mold patterning allows the orientation of ceramic nanoparticles into parallel lines within the composite scaffold. The ability of the produced films to host and address cell growth is evaluated after 1, 3, and 6 days of culturing with murine derived cardiac and mesenchymal stem cells (CSCs and MSCs), and compared with PLGA films without ceramics and loaded with nanostructured TiO2. Aligned cell growth is observed only for scaffolds that incorporate oriented ceramic nanoparticles, attributed to the nanoceramic ability to modulate the roughness pitch, thus improving cell sensitivity towards the host surface features. Better CSC and MSC proliferative activity is observed for CeO2 composites with respect to either TiO2 -added or unfilled PLGA films. This evidence may be related to the nanostructured CeO2 antioxidative properties. [source]

Age Dependence of the Human Skeletal Muscle Stem Cell in Forming Muscle Tissue

ARTIFICIAL ORGANS, Issue 3 2006
Ralf Schäfer
Abstract:, Human skeletal muscle stem cells from healthy donors aged 2,82 years (n = 13) and from three children suffering from Duchenne Muscular Dystrophy (DMD) were implanted into soleus muscles of immunoincompetent mice and were also expanded in vitro until senescence. Growth of implanted cells was quantified by structural features and by the amount of human DNA present in a muscle. Proliferative capacity in vitro and in vivo was inversely related to age of the donor. In vitro, a decline of about two mean population doublings (MPDs) per 10 years of donor's age was observed. Muscle stem cells from DMD children were prematurely aged. In general, cell preparations with low or decreasing content in desmin-positive cells produced more MPDs than age-matched high-desmin preparations and upon implantation more human DNA and more nonmyogenic than myogenic tissue. Thus, a "Desmin Factor" was derived which predicts "quality" of the human muscle tissue growing in vivo. This factor may serve as a prognostic tool. [source]

Adipogenic Differentiation of Human Adipose Tissue,Derived Stem Cells Obtained from Cryopreserved Adipose Aspirates

DERMATOLOGIC SURGERY, Issue 7 2010
JUNG EUN LEE MS
BACKGROUND Although frozen adipose tissue is frequently used for soft tissue augmentation, the viability of frozen fat remains a controversy. The cryopreservation of adipose tissue is important for the future use of adipose-derived stem cells (ASCs) and adipocytes. OBJECTIVE To determine whether optimal cryopreservation techniques with regard to the addition of cryopreservative agents and preservation temperature is essential for the long-term storage of adipose tissue and whether ASCs from cryopreserved adipose aspirates are reliable for use in adipogenic differentiation. MATERIALS AND METHODS Adipose tissue was frozen directly or with cryoprotectant at ,20°C or ,80°C for 1 year. The viability of adipose aspirates and the differentiation of ASCs isolated from adipose tissue were evaluated. RESULTS The viability of adipose aspirates frozen with dimethyl sulfoxide at ,80°C was approximately 87% after 2 months of storage. Moreover, ASCs from adipose tissue stored with cryoprotectant survived successfully for 1 year and differentiated into adipocytes, although ASCs were not detected in the directly frozen adipose tissue. CONCLUSION Adipose tissue cryopreserved with cryoprotectant and stored at optimal temperature might prove to be a reliable source of human ASCs and adipocytes. The authors have indicated no significant interest with commercial supporters. [source]

Dual-Growth-Factor-Releasing PCL Scaffolds for Chondrogenesis of Adipose-Tissue-Derived Mesenchymal Stem Cells,

ADVANCED ENGINEERING MATERIALS, Issue 1-2 2010
Sung Mook Lim
Polycaprolactone/Pluronic F127 porous scaffolds are prepared using a modified melt-molding particulate-leaching method. The scaffolds are highly porous (about 90% porosity) and have open-cellular pore structures. Growth factors (TGF- ,2, BMP-7 or dual TGF- ,2/BMP-7) can be easily immobilized on the pore surfaces of the PCL/F127 scaffolds via binding with heparin. The growth-factor-immobilized scaffolds can induce the chondrogenesis of ATMSCs seeded onto them. Using TGF-,2 and BMP-7 growth factors together leads to a better chondrogenic differentiation behavior than using single-growth-factor immobilized scaffolds. [source]

A New Approach for Adipose Tissue Regeneration Based on Human Mesenchymal Stem Cells in Contact to Hydrogels,an In Vitro Study,

ADVANCED ENGINEERING MATERIALS, Issue 10 2009
Kirsten Peters
In this study an approach for adipose tissue regeneration based on human mesenchymal stem cells and hydrogels as supporting matrix was evaluated. The gelatin-based hydrogels developed in this study were cytocompatible and stem cell adhesion onto hydrogel surfaces was higher as compared to tissue culture polystyrene. Furthermore, the adipogenic differentiation degree was increased. These results are promising for future applications of hydrogels in adipose tissue regeneration strategies. [source]

Revelationary biology: A review of The Second Tree: Stem Cells, Clones, Chimeras, and Quests for Immortality, by Elaine Dewar

EVOLUTION AND DEVELOPMENT, Issue 5 2005
Yolanda P. Cruz
First page of article [source]

Embryonic Stem Cells and Gene Targeting

EXPERIMENTAL PHYSIOLOGY, Issue 6 2000
Birgit Ledermann
The development of gene targeting technology, the exchange of an endogenous allele of a target gene for a mutated copy via homologous recombination, and the application of this technique to murine embryonic stem cells has made it possible to alter the germ-line of mice in a predetermined way. Gene targeting has enabled researchers to generate mouse strains with defined mutations in their genome allowing the analysis of gene function in vivo. This review presents the essential tools and methodologies used for gene targeting that have been developed over the past decade. Special emphasis has been laid on the available embryonic stem cell lines and the importance of the genetic background. Also, the state-of-the art of gene targeting approaches in species other than mice will be discussed. [source]

Labeling of Adipose-Derived Stem Cells by Oleic-Acid-Modified Magnetic Nanoparticles

ADVANCED FUNCTIONAL MATERIALS, Issue 8 2009
Lian Cen
Abstract The in vivo tracking of adipose derived stem cells (ASCs) is of essential concern when they are used as seed cells in tissue engineering. This study explores the feasibility of using magnetic nanoparticles (MNs), a type of contrast agents in magnetic resonance imaging (MRI), to label ASCs such that the labeled ASCs could be tracked in vivo by MRI non-invasively and repeatedly. To do this, MNs of <10,nm surface-coated with oleic acid are synthesized via a high-temperature solution-phase reaction. Cytotoxicity of the as-synthesized MNs at concentrations up to 0.1,mg,mL,1 on 104,cells,mL,1 ASCs is evaluated by LDH release. Since only minor cytotoxicity is detected, the effects of the labeling technique on cellular behaviors and uptake by labeled cells are investigated. Cell proliferation and differentiation with and without MNs are compared. The results show that proliferation of ASCs (104,cells,mL,1) labeled by MNs (0.05,mg,mL,1) is significantly enhanced and dependent on the labeling time. The MNs are located in the vesicles within cytoplasm of ASCs. The cellular uptake reaches as high as ,180,pg/cell. Nevertheless, the labeled ASCs still maintained adipogenic and osteogenic differentiation. Hence, the feasibility of labeling ASCs by oleic acid coated MNs is ascertained and it was better to label the cells during their quiescent stage. The labeled ASCs can also be in vivo detected by MRI in a subcutaneous model in vivo. Further MRI tracking of the labeled ASCs in long-term follow-up would thus follow this current study. [source]

Mapping the Interactions among Biomaterials, Adsorbed Proteins, and Human Embryonic Stem Cells

ADVANCED MATERIALS, Issue 27 2009
Ying Mei
An integrated high-throughput polymer synthesis and rapid material/protein/cell interaction assays were developed to optimize stem cell microenvironments. Microarrayed polymers were synthesized and studied for the ability to support the growth of partially differentiated human embryonic stem cells. In parallel, a programmed laser scanning cytometry system was developed to allow for rapid quantification of cell material interaction. [source]

Carbon Nanotube Monolayer Patterns for Directed Growth of Mesenchymal Stem Cells,

ADVANCED MATERIALS, Issue 18 2007
Y. Park
Carbon nanotube (CNT) monolayer patterns are utilized to control the growth of mesenchymal stem cells (MSCs) (see figure). MSCs exhibit preferential growth on CNT patterns, suggesting that the CNT monolayer does not have a harmful effect on the MSCs. Furthermore, the growth of MSCs on swCNT patterns between electrodes is demonstrated. These results show that CNT patterns have enormous potential as a new platform for basic research and applications using stem cells. [source]

Restoration of Bone Mass and Strength in Glucocorticoid-Treated Mice by Systemic Transplantation of CXCR4 and Cbfa-1 Co-Expressing Mesenchymal Stem Cells,

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 5 2009
Chun-Yang Lien
Abstract Transplantation of gene-modified mesenchymal stem cells (MSCs) in animals for bone regeneration therapy has been evaluated extensively in recent years. However, increased endosteal bone formation by intravenous injection of MSCs ectopically expressing a foreign gene has not yet been shown. Aside from the clearance by lung and other tissues, the surface compositions of MSCs may not favor their bone marrow (BM) migration and engraftment. To overcome these hurdles, a gene encoding the chemokine receptor largely responsible for stromal-derived factor-1 (SDF-1)-mediated BM homing and engraftment of hematopoietic stem cells (HSCs), CXCR4, was transduced into mouse C3H10T1/2 cells by adenovirus infection. A dose-dependent increase of CXCR4 surface expression with a parallel enhanced chemotaxis toward SDF-1 in these cells after virus infection was clearly observed. Higher BM retention and homing of CXCR4-expressing MSCs were also found after they were transplanted by intramedullary and tail vein injections, respectively, into immunocompetent C3H/HeN mice. Interestingly, a full recovery of bone mass and a partial restoration of bone formation in glucocorticoid-induced osteoporotic mice were observed 4 wk after a single intravenous infusion of one million CXCR4-expressing C3H10T1/2 cells. In the meantime, complete recovery of bone stiffness and strength in these animals was consistently detected only after a systemic transplantation of CXCR4 and Cbfa-1 co-transduced MSCs. To our knowledge, this is the first report to show unequivocally the feasibility of ameliorating glucocorticoid-induced osteoporosis by systemic transplantation of genetically manipulated MSCs. [source]

Activation of Sirt1 Decreases Adipocyte Formation During Osteoblast Differentiation of Mesenchymal Stem Cells,

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 7 2006
Carl-Magnus Bäckesjö PhD
Abstract In vitro, mesenchymal stem cells differentiate to osteoblasts when exposed to bone-inducing medium. However, adipocytes are also formed. We showed that activation of the nuclear protein deacetylase Sirt1 reduces adipocyte formation and promotes osteoblast differentiation. Introduction: Mesenchymal stem cells (MSCs) can differentiate into osteoblasts, adipocytes, chondrocytes, and myoblasts. It has been suggested that a reciprocal relationship exists between the differentiation of MSCs into osteoblasts and adipocytes. Peroxisome proliferator-activated receptor ,2 (PPAR,2) is a key element for the differentiation into adipocytes. Activation of Sirt1 has recently been shown to decrease adipocyte development from preadipocytes through inhibition of PPAR,2. Materials and Methods: We used the mouse mesenchymal cell line C3H10T1/2 and primary rat bone marrow cells cultured in osteoblast differentiation medium with or without reagents affecting Sirt1 activity. Adipocyte levels were analyzed by light microscopy and flow cytometry (FACS) after staining with Oil red O and Nile red, respectively. Osteoblast and adipocyte markers were studied with quantitative real-time PCR. Mineralization in cultures of primary rat bone marrow stromal cells was studied by von Kossa and alizarin red staining. Results: We found that Sirt1 is expressed in the mesenchymal cell line C3H10T1/2. Treatment with the plant polyphenol resveratrol as well as isonicotinamide, both of which activate Sirt1, blocked adipocyte development and increased the expression of osteoblast markers. Nicotinamide, which inhibits Sirt1, increased adipocyte number and increased expression of adipocyte markers. Furthermore, activation of Sirt1 prevented the increase in adipocytes caused by the PPAR,-agonist troglitazone. Finally, activation of Sirt1 in rat primary bone marrow stromal cells increased expression of osteoblast markers and also mineralization. Conclusions: In this study, we targeted Sirt1 to control adipocyte development during differentiation of MSCs into osteoblasts. The finding that resveratrol and isonicotinamide markedly inhibited adipocyte and promoted osteoblast differentiation may be relevant in the search for new treatment regimens of osteoporosis but also important for the evolving field of cell-based tissue engineering. [source]

Strategies for Directing the Differentiation of Stem Cells Into the Osteogenic Lineage In Vitro,

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 9 2004
Boon Chin Heng
Abstract A major area in regenerative medicine is the application of stem cells in bone reconstruction and bone tissue engineering. This will require well-defined and efficient protocols for directing the differentiation of stem cells into the osteogenic lineage, followed by their selective purification and proliferation in vitro. The development of such protocols would reduce the likelihood of spontaneous differentiation of stem cells into divergent lineages on transplantation, as well as reduce the risk of teratoma formation in the case of embryonic stem cells. Additionally, such protocols could provide useful in vitro models for studying osteogenesis and bone development, and facilitate the genetic manipulation of stem cells for therapeutic applications. The development of pharmokinetic and cytotoxicity/genotoxicity screening tests for bone-related biomaterials and drugs could also use protocols developed for the osteogenic differentiation of stem cells. This review critically examines the various strategies that could be used to direct the differentiation of stem cells into the osteogenic lineage in vitro. [source]

Cancer stem cells in leukemia, recent advances

JOURNAL OF CELLULAR PHYSIOLOGY, Issue 2 2007
Gang-Ming Zou
The history of stem cell research was started in the early 1900s in Europe where the researcher realized that various types of blood cells came from a particular "stem cells." However, it was not until 1963 that the first quantitative description of the self-renewal activities of transplanted mouse bone marrow cells were documented by Canadian scientist Ernest A McCulloch and James E Till in Toronto. The concept of cancer stem cells has been used over 50 years ago; whereas the strong evidence for the existence of a Cancer Stem Cells was obtained recently. Consequently, there is increasing attention in recent year about cancer stem cells. The findings from recent studies support the concept that stem cells are integral to the development of several forms of human cancer. Changes in stem cell behavior can contribute to tumor formation. Leukemia is a cancer of blood-forming tissue, including the bone marrow and lymphatic system. Leukemic stem cells represent the cancer stem cells in the leukemia. In this review, we summarize the recent advance in the study of leukemic stem cells. J. Cell. Physiol. 213: 440,444, 2007. © 2007 Wiley-Liss, Inc. [source]

Welcome to the new section of the Editorial Board of Aging Cell: Stem Cells in Aging

AGING CELL, Issue 4 2005
Tim Cowen
No abstract is available for this article. [source]

Poster Sessions BP06: Neurogenesis, Stem Cells and Apoptosis

JOURNAL OF NEUROCHEMISTRY, Issue 2002
J. Qiu
The NF-,B transcription factor regulates bcl-x gene expression, which may determine hypoxia-induced neuronal apoptosis. We examined hypoxia-induced NF-,B and Bcl-xL changes in rat hippocampus. We showed differential hypoxia-induced NF-,B binding to the bcl-x promoter CS4 and IgG,,B enhancer sequences by rat hippocampal nuclear extracts. The differential NF-,B binding to these two promoter sequences was also determined in a contused spinal cord injury model and in vitro studies with LPS-treated Hela cells. There was tissue-, gene promoter-specific and time-dependent regulation of bcl-x gene expression by NF-,B in support of the hypothesis that NF-,B has tissue- and gene-specific regulatory effects and that these effects may account for both the pro- and anti-apoptotic roles assigned to NF-,B in different apoptotic processes. We applied ,decoy' oligonucleotides with sequences specific to different promoters to the rat hippocampus and measured ,decoy' inhibitory effects on nuclear NF-,B binding. The IgG-,B enhancer sequence ,decoy' showed stronger inhibition on nuclear NF-,B c-Rel/p50 binding to the bcl-x gene promoter CS4 sequence when compared to NF-,B p50/p50 binding to the same sequence. This result suggests that the ,decoy' approach has the potential to selectively manipulate NF-,B regulation of gene expression in response to hypoxia. Acknowledgements:, Supported by NINDS NS-39161, Shriner Grant 8710 and a Grant from the Sealy Center on Aging to J. Qiu. [source]

Symposium 10: Differentiation Plasticity of Stem Cells

JOURNAL OF NEUROCHEMISTRY, Issue 2002
S. S. Liour
The major role of radial glial cells in neuronal development is to provide support and guidance for neuronal migration. In vitro, neurons, astrocytes and oligodendrocytes have also been generated from neural stem cells and embryonic stem cells, but the generation of radial glial cells in vitro has not yet been reported. Since radial glial cells can lead to neurons and astrocytes during brain development, neurogenesis and gliogenesis of stem cells in vitro may at least in part also utilize the same mechanisms. To test this hypothesis, we utilized five different clones of embryonic (ES) and embryonal carcinoma (EC) stem cell lines to investigate the differentiation of radial glial cells during in vitro neural differentiation. Here, we demonstrate that radial glial cells can be generated from ES/EC cell lines. These ES/EC cell-derived radial glial cells are similar in morphology to radial glial cells in vivo. They also express several cytoskeletal markers that are characteristics of radial glial cells in vivo. The processes of these in vitro -generated radial glial cells are organized into scaffolds that appear to support the migration of newly generated neurons in culture. Like radial glial cells in vivo, they appear to differentiate subsequently into astrocytes. Differentiation of radial glial cells may be a common pathway during in vitro neural differentiation of ES cells. This novel in vitro model system may facilitate the investigation of regulation of radial glial cell differentiation and its biological function. Acknowledgements:, Supported by USPHS Grant NS11853 and a grant from the Children's Medical Research Foundation. [source]

The Neuropeptide Pituitary Adenylate Cyclase-Activating Polypeptide Exerts Anti-Apoptotic and Differentiating Effects during Neurogenesis: Focus on Cerebellar Granule Neurones and Embryonic Stem Cells

JOURNAL OF NEUROENDOCRINOLOGY, Issue 5 2007
A. Falluel-Morel
Pituitary adenylate cyclase-activating polypeptide (PACAP) was originally isolated from ovine hypothalamus on the basis of its hypophysiotrophic activity. It has subsequently been shown that PACAP and its receptors are widely distributed in the central nervous system of adult mammals, indicating that PACAP may act as a neurotransmitter and/or neuromodulator. It has also been found that PACAP and its receptors are expressed in germinative neuroepithelia, suggesting that PACAP could be involved in neurogenesis. There is now compelling evidence that PACAP exerts neurotrophic activities in the developing cerebellum and in embryonic stem (ES) cells. In particular, the presence of PACAP receptors has been demonstrated in the granule layer of the immature cerebellar cortex, and PACAP has been shown to promote survival, inhibit migration and activate neurite outgrowth of granule cell precursors. In cerebellar neuroblasts, PACAP is a potent inhibitor of the mitochondrial apoptotic pathway through activation of the MAPkinase extracellular regulated kinase. ES cells and embryoid bodies (EB) also express PACAP receptors and PACAP facilitates neuronal orientation and induces the appearance of an electrophysiological activity. Taken together, the anti-apoptotic and pro-differentiating effects of PACAP characterised in cerebellar neuroblasts as well as ES and EB cells indicate that PACAP acts not only as a neurohormone and a neurotransmitter, but also as a growth factor. [source]

Ethanol Alters the Osteogenic Differentiation of Amniotic Fluid-Derived Stem Cells

ALCOHOLISM, Issue 10 2010
Jennifer A. Hipp
Background:, Fetal alcohol spectrum disorder (FASD) is a set of developmental defects caused by prenatal alcohol exposure. Clinical manifestations of FASD are highly variable and include mental retardation and developmental defects of the heart, kidney, muscle, skeleton, and craniofacial structures. Specific effects of ethanol on fetal cells include induction of apoptosis as well as inhibition of proliferation, differentiation, and migration. This complex set of responses suggests that a bioinformatics approach could clarify some of the pathways involved in these responses. Methods:, In this study, the responses of fetal stem cells derived from the amniotic fluid (AFSCs) to treatment with ethanol have been examined. Large-scale transcriptome analysis of ethanol-treated AFSCs indicates that genes involved in skeletal development and ossification are up-regulated in these cells. Therefore, the effect of ethanol on osteogenic differentiation of AFSCs was studied. Results:, Exposure to ethanol during the first 48 hours of an osteogenic differentiation protocol increased in vitro calcium deposition by AFSCs and increased alkaline phosphatase activity. In contrast, ethanol treatment later in the differentiation protocol (day 8) had no significant effect on the activity of alkaline phosphatase. Conclusions:, These results suggest that transient exposure of AFSCs to ethanol during early differentiation enhances osteogenic differentiation of the cells. [source]

Effects of Ethanol on Mouse Embryonic Stem Cells

ALCOHOLISM, Issue 12 2009
Alla Arzumanyan
Background:, Fetal alcohol syndrome (FAS) reflects a constellation of congenital abnormalities caused by excess maternal consumption of alcohol. It is likely that interference with embryonic development plays a role in the pathogenesis of the disorder. Ethanol-induced apoptosis has been suggested as a causal factor in the genesis of FAS. Mouse embryonic stem (mES) cells are pluripotent cells that differentiate in vitro to cell aggregates termed embryoid bodies (EBs), wherein differentiation capacity and gene expression profile are similar to those of the early embryo. Methods:, To investigate the effects of ethanol during differentiation, mES cells were cultured on a gelatin surface in the presence of leukemia inhibitory factor which maintains adherent undifferentiated cells or in suspension to promote formation of EBs. All cells were treated (1,6 days) with 80 mM ethanol. The pluripotency and differentiation of mES cells were evaluated by western blotting of stage-specific embryonic antigen (SSEA-1), transcription factors Oct-3/4, Sox-2, and Nanog, using alkaline phosphatase staining. Apoptosis (early to late stages) was assessed by fluorescence-activated cell sorting using TdT-mediated biotin,dUTP nick-end labelling assay and fluorescein isothiocyanate-Annexin V/propidium iodide staining. Results:, Ethanol increased apoptosis during in vitro differentiation of mES cells to EBs, whereas undifferentiated cells were not affected. Ethanol exposure also interfered with pluripotency marker patterns causing an upregulation of SSEA-1 under self-renewal conditions. In EBs, ethanol delayed the downregulation of SSEA-1 and affected the regulation of transcription factors during differentiation. Conclusion:, Our findings suggest that ethanol may contribute to the pathogenesis of FAS by triggering apoptotic pathways during differentiation of embryonic stem cells and deregulating early stages of embryogenesis. [source]

Adipogenic Effect of Alcohol on Human Bone Marrow-Derived Mesenchymal Stem Cells

ALCOHOLISM, Issue 7 2004
Frederick H. Wezeman
Background: In addition to a decrease in bone mass in alcoholics their osteopenic skeletons show an increase in bone marrow adiposity. Human bone marrow mesenchymal stem cells (hMSC) in vivo differentiate into several phenotypes including osteogenic and adipogenic cells, both of which remain as resident populations of bone marrow. In vitro, the lineage commitment and differentiation of hMSC toward the adipogenic pathway can be promoted by alcohol. Methods: Human male and female mesenchymal stem cells from joint replacement surgery were cultured. Cells were grouped as: 1) Control (no additions to the culture medium), 2) EtOH (50 mm alcohol added to the culture medium), 3) OS (osteogenic inducers added to the culture medium), and 4) OS + EtOH (osteogenic inducers and 50 mm alcohol added to the culture medium). Cultures stained with Nile Red confirmed the development of differentiated adipocytes. Population analysis was performed using fluorescence-activated cell sorting. Gene expression of early, middle, late, and terminal differentiation stage markers (PPAR),2, lipoprotein lipase, adipsin, leptin, and adipocyte P2 (aP2)] was studied by Northern hybridization, and protein synthesis of aP2 was determined by Western analysis. Results: Nile red staining confirmed increased adipocyte development 10 days after the onset of treatment with 50 mm alcohol and osteogenic induction. By day 21 the number of adipocytes increased to 13.6% of the total population. Alcohol up-regulated the gene expression of PPAR,2 whereas no up-regulation was observed for the other genes. Protein production of aP2 was significantly increased in hMSC cells by culture in the presence of alcohol. Conclusions: The data suggest that alcohol's adipogenic effect on cultured hMSC is through up-regulation of PPAR,2 at the point of lineage commitment as well as through enhancement of lipid transport and storage through increased aP2 synthesis. The alcohol-induced expression and synthesis changes account for the increased Nile red staining of cultured hMSC. [source]

Alcohol-Induced Neurodegeneration: When, Where and Why?

ALCOHOLISM, Issue 2 2004
Fulton T. Crews
Abstract: This manuscript reviews the proceedings of a symposium organized by Drs. Antonio Noronha and Fulton Crews presented at the 2003 Research Society on Alcoholism meeting. The purpose of the symposium was to examine recent findings on when alcohol induced brain damage occurs, e.g., during intoxication and/or during alcohol withdrawal. Further studies investigate specific brain regions (where) and the mechanisms (why) of alcoholic neurodegeneration. The presentations were (1) Characterization of Synaptic Loss in Cerebella of Mature and Senescent Rats after Lengthy Chronic Ethanol Consumption, (2) Ethanol Withdrawal Both Causes Neurotoxicity and Inhibits Neuronal Recovery Processes in Rat Organotypic Hippocampal Cultures, (3) Binge Drinking-Induced Brain Damage: Genetic and Age Related Effects, (4) Binge Ethanol-Induced Brain Damage: Involvement of Edema, Arachidonic Acid and Tissue Necrosis Factor , (TNF,), and (5) Cyclic AMP Cascade, Stem Cells and Ethanol. Taken together these studies suggest that alcoholic neurodegeneration occurs through multiple mechanisms and in multiple brain regions both during intoxication and withdrawal. [source]

Neural Stem Cells and Alcohol

ALCOHOLISM, Issue 2 2003
F. T. Crews
This article summarizes the proceedings of a symposium held at the 2002 Research Society on Alcoholism Meeting in San Francisco, California. The aim of this symposium was to review research on the effects of ethanol on neural stems cells and neurogenesis. Ethanol is known to alter neurogenesis during development; however, recent studies indicate that the brain forms new neurons from stem cells throughout life. Furthermore, stem cells can be transplanted into the brain, creating exciting new possibilities to study brain function. The symposium covered these research areas. Dr. Michael W. Miller reviewed knowledge on the effects of ethanol on stem cell proliferation and differentiation during development. Dr. Wu Ma described studies in culture indicating that (1) neural stem cells express functional muscarinic acetylcholine receptors (mAchR), (2) mAchR-mediated proliferation involves Ca2+ signaling and mitogen-activated protein kinase phosphorylation, and (3) phosphoinositol-3 kinase is a downstream effector for mAchR-mediated cell proliferation via activation of Akt. Drs. Kim Nixon and Fulton T. Crews followed with in vivo studies on ethanol's effects on adult neural stem cell proliferation and differentiation. Dr. W. Michael Zawada described studies directed at dopamine neuron cell transplants into mammalian central nervous system. These studies clearly establish that ethanol has significant effects on stem cells. [source]

A Micro/Nanoscale Surface Mechanical Study on Morpho-Functional Changes in Multilineage-Differentiated Human Mesenchymal Stem Cells

MACROMOLECULAR BIOSCIENCE, Issue 5 2007
Serena Danti
Abstract In recent years MSCs have become a very attractive tool in tissue engineering and regenerative medicine because of their ability to be committed along several lineages through chemical or physical stimuli. Nevertheless their therapeutic potential and plasticity are not yet totally understood. This report describes the use of AFM together with conventional microscopies to obtain mechanical information on cell surfaces and deposited extra cellular matrix molecules, after inducing the differentiation of human MSCs towards three typical mesoderm phenotypes. The aim is to correlate morphological, functional, and mechanical aspects of human MSCs to obtain a deeper understanding of their great potential. [source]

Polypyrrole Thin Films Formed by Admicellar Polymerization Support the Osteogenic Differentiation of Mesenchymal Stem Cells

MACROMOLECULAR BIOSCIENCE, Issue 8 2004
Harold Castano
Abstract Summary: The objective of this study was to evaluate the attachment, proliferation, and differentiation of rat mesenchymal stem cells (MSC) toward the osteoblastic phenotype seeded on polypyrrole (PPy) thin films made by admicellar polymerization. Three different concentrations of pyrrole (Py) monomer (20, 35, and 50,×,10,3M) were used with the PPy films deposited on tissue culture polystyrene dishes (TCP). Regular TCP dishes and PPy polymerized on TCP by chemical polymerization without surfactant using 5,×,10,3M Py, were used as controls. Rat MSC were seeded on these surfaces and cultured for up to 20 d in osteogenic media. Surface topography was characterized by atomic force microscopy, X-ray photoelectron spectroscopy, and static contact angle. Cell attachment, proliferation, alkaline phosphatase (ALP) activity, and calcium content were measured to evaluate the ability of MSC to adhere and differentiate on PPy-coated TCP. Increased monomer concentrations resulted in PPy films of increased thickness and surface roughness. PPy films generated by different monomer concentrations induced drastically different cellular events. A wide spectrum of cell attachment characteristics (from excellent cell attachment to the complete inability to adhere) were obtained by varying the monomer concentration from 20 m to 50,×,10,3M. In particular the 20,×,10,3M PPy thin films demonstrated superior induction of MSC osteogenicity, which was comparable to standard TCP dishes, unlike PPy films of similar thickness prepared by chemical polymerization without surfactant. Adhesion of mesenchymal stem cells on tissue culture plates (TCP) coated with polypyrrole thin films made by admicellar polymerization. [source]

FGF Ligand Family mRNA Expression Profile for Mouse Preimplantation Embryos, Early Gestation Human Placenta, and Mouse Trophoblast Stem Cells

MOLECULAR REPRODUCTION & DEVELOPMENT, Issue 5 2006
W. Zhong
Abstract Signaling by fibroblast growth factor (FGF) is essential is for trophoblast stem (TS) cells and preimplantation embryos. FGF4 provides essential signaling, but the expression of the complete set of 23 FGF family members has not been analyzed. Here, semi-quantitative RT-PCR and microarray analyses were used to define expression of all FGF ligand mRNA. RT-PCR was done for developmentally important FGF subfamilies, FGF10/FGF22 and FGF8/FGF17/FGF18 as well as FGF11. FGF4 and FGF18 are detected at highest levels by RT-PCR and microarrays. FGF10 was detected at low levels in both assays. FGF11 was detected at moderate levels by microarray, but not by RT-PCR. FGF17 was detected at low levels by array and moderate levels by RT-PCR. FGF8 and FGF22 were detected by RT-PCR, but not by microarrays during late cleavage divisions. FGF8, FGF5, and FGF9 were detected in the oocyte by microarray. FGF2, FGF3, and FGF7 were not detected by RT-PCR or microarrays and FGF13, FGF14, and FGF23 were not detected by microarray. Since a major role of FGF is to maintain TS cells, we tested human and mouse placental cell lines and early gestation human placenta for expression of FGF ligands. Expression in mouse TS cells was compared with preimplantation embryos, and human placental cell line expression was compared with human placenta, to infer which ligands are expressed in placental lineage vs. other cell lineages. The data suggest that human and mouse placenta share FGF18 and its high expression suggests preimplantation and early placental function. Mol. Reprod. Dev. © 2006 Wiley-Liss, Inc. [source]

Will Embryonic Stem Cells Change Health Policy?

THE JOURNAL OF LAW, MEDICINE & ETHICS, Issue 2 2010
William M. Sage
Embryonic stem cells are actively debated in political and public policy arenas. However, the connections between stem cell innovation and overall health care policy are seldom elucidated. As with many controversial aspects of medical care, the stem cell debate bridges to a variety of social conversations beyond abortion. Some issues, such as translational medicine, commercialization, patient and public safety, health care spending, physician practice, and access to insurance and health care services, are core health policy concerns. Other issues, such as economic development, technologic progress, fiscal politics, and tort reform, are only indirectly related to the health care system but are frequently seen through a health care lens. These connections will help determine whether the stem cell debate reaches a resolution, and what that resolution might be. [source]