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Anticoagulants on yield of bone marrow-derived mononuclear cells harvested from dogs Ciência Rural
Brito,Harald Fernando Vicente de; Fernandes-Filho,Tobias; Brandão,Fernanda de Lima; Montaño,Patrícia Yukiko; Laskoski,Luciane Maria; Locatelli-Dittrich,Rosangela.
Cell therapy with bone marrow-derived mononuclear cells is an alternative to therapy with mesenchymal stem cell cultures. The aim of the present research was the comparison of the yield of bone marrow-derived mononuclear cells harvested from dogs with two different anticoagulants. Bone marrow was harvested from the iliac crest of five healthy dogs aged between 15 and 30 months, and the effect of two anticoagulant solutions, CPDA-1 (citrate phosphate dextrose adenine-1) and heparin, on the isolation of mononuclear cells was compared. Mononuclear cells were isolated in a density gradient and stained for CD9 and CD44 for characterization by flow cytometry. Means were compared using Student's paired t-test. Samples harvested with CPDA-1 yielded an average of...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mesenchymal stem cells; Multipotent stem cell; Immunophenotyping.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782015001102044
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bFGF promotes adipocyte differentiation in human mesenchymal stem cells derived from embryonic stem cells Genet. Mol. Biol.
Song,Xinghui; Li,Yanwei; Chen,Xiao; Yin,Guoli; Huang,Qiong; Chen,Yingying; Xu,Guowei; Wang,Linlin.
In this work we describe the establishment of mesenchymal stem cells (MSCs) derived from embryonic stem cells (ESCs) and the role of bFGF in adipocyte differentiation. The totipotency of ESCs and MSCs was assessed by immunofluorescence staining and RT-PCR of totipotency factors. MSCs were successfully used to induce osteoblasts, chondrocytes and adipocytes. MSCs that differentiated into adipocytes were stimulated with and without bFGF. The OD/DNA (optical density/content of total DNA) and expression levels of the specific adipocyte genes PPARγ2 (peroxisome proliferator activated receptor γ2) and C/EBPs were higher in bFGF cells. Embryonic bodies had a higher adipocyte level compared with cells cultured in plates. These findings indicate that bFGF promotes...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Embryonic stem cells; Mesenchymal stem cells; Fibroblast growth factor; Adipocyte differentiation.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572014000100019
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CTLA4 enhances the osteogenic differentiation of allogeneic human mesenchymal stem cells in a model of immune activation BJMBR
Dai,F.; Zhang,F.; Sun,D.; Zhang,Z.H.; Dong,S.W.; Xu,J.Z..
Allogeneic mesenchymal stem cells (allo-MSCs) have recently garnered increasing interest for their broad clinical therapy applications. Despite this, many studies have shown that allo-MSCs are associated with a high rate of graft rejection unless immunosuppressive therapy is administered to control allo-immune responses. Cytotoxic T-lymphocyte-associated protein 4 (CTLA4) is a co-inhibitory molecule expressed on T cells that mediates the inhibition of T-cell function. Here, we investigated the osteogenic differentiation potency of allo-MSCs in an activated immune system that mimics the in vivo allo-MSC grafting microenvironment and explored the immunomodulatory role of the helper T cell receptor CTLA4 in this process. We found that MSC osteogenic...
Tipo: Info:eu-repo/semantics/article Palavras-chave: CTLA4; Mesenchymal stem cells; Immunomodulatory; Bone tissue engineering.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2015000700629
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Effect of conditioned medium of mesenchymal stem cells on the in vitro maturation and subsequent development of mouse oocyte BJMBR
Ling,B.; Feng,D.Q.; Zhou,Y.; Gao,T.; Wei,H.M.; Tian,Z.G..
Mesenchymal stem cells (MSCs) secrete a variety of cytokines and growth factors in addition to self-renewal and multiple forms of differentiation. Some of these secreted bioactive factors could improve meiotic maturation in vitro and subsequent embryo developmental potential. The aim of the present study was to determine whether in vitro maturation (IVM) of mouse oocyte with or without cumulus cells could be improved by contact with conditioned medium (CM) of MSCs as well as the efficiency of CM to support follicular growth and oocyte maturation in the ovarian organ of mice cultured on soft agar. The developmental potential of matured oocyte was assessed by blastocyst formation after in vitro fertilization (IVF). Germinal vesicle stage oocytes with or...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mesenchymal stem cells; In vitro maturation; Follicular development; Mouse oocyte development.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2008001100006
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Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development BJMBR
Feng,D.Q.; Zhou,Y.; Ling,B.; Gao,T.; Shi,Y.Y.; Wei,H.M.; Tian,Z.G..
Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca²+]i), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mesenchymal stem cells; Oocyte activation; Parthenogenesis; Calcium; Cytochalasin B.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2009000600006
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Fibrin biopolymer as scaffold candidate to treat bone defects in rats J. Venom. Anim. Toxins incl. Trop. Dis.
Cassaro,Claudia Vilalva; Justulin Jr.,Luis Antonio; Lima,Patrícia Rodrigues de; Golim,Marjorie de Assis; Biscola,Natália Perussi; Castro,Mateus Vidigal de; Oliveira,Alexandre Leite Rodrigues de; Doiche,Danuta Pulz; Pereira,Elenize Jamas; Ferreira Jr.,Rui Seabra; Barraviera,Benedito.
ABSTRACT Background: Bone tissue repair remains a challenge in tissue engineering. Currently, new materials are being applied and often integrated with live cells and biological scaffolds. The fibrin biopolymer (FBP) proposed in this study has hemostatic, sealant, adhesive, scaffolding and drug-delivery properties. The regenerative potential of an association of FBP, biphasic calcium phosphate (BCP) and mesenchymal stem cells (MSCs) was evaluated in defects of rat femurs. Methods: Adult male Wistar rats were submitted to a 5-mm defect in the femur. This was filled with the following materials and/or associations: BPC; FBP and BCP; FBP and MSCs; and BCP, FBP and MSCs. Bone defect without filling was defined as the control group. Thirty and sixty days...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bone regeneration; Biomaterials; Fibrin sealant; Fibrin biopolymer; Biphasic calcium phosphate; Mesenchymal stem cells.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992019000100320
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Human mesenchymal stem cells are resistant to cytotoxic and genotoxic effects of cisplatin in vitro Genet. Mol. Biol.
Bellagamba,Bruno Corrêa; Abreu,Bianca Regina Ribas de; Grivicich,Ivana; Markarian,Carolina Franke; Chem,Eduardo; Camassola,Melissa; Nardi,Nance Beyer; Dihl,Rafael Rodrigues.
Abstract Mesenchymal stem cells (MSCs) are known for their important properties involving multilineage differentiation potential., trophic factor secretion and localization along various organs and tissues. On the dark side, MSCs play a distinguished role in tumor microenvironments by differentiating into tumor-associated fibroblasts or supporting tumor growth via distinct mechanisms. Cisplatin (CIS) is a drug widely applied in the treatment of a large number of cancers and is known for its cytotoxic and genotoxic effects, both in vitro and in vivo. Here we assessed the effects of CIS on MSCs and the ovarian cancer cell line OVCAR-3, by MTT and comet assays. Our results demonstrated the resistance of MSCs to cell death and DNA damage induction by CIS,...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Mesenchymal stem cells; Cisplatin; MTT; Comet assay; OVCAR-3 cells.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572016000100129
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Immunomodulatory effect of mesenchymal stem cells BJMBR
Herrero,C.; Pérez-Simón,J.A..
Mesenchymal stem cells (MSC) are multipotential nonhematopoietic progenitor cells capable of differentiating into multiple mesenchymal tissues. MSC are able to reconstitute the functional human hematopoietic microenvironment and promote engraftment of hematopoietic stem cells. MSC constitutively express low levels of major histocompatibility complex-I molecules and do not express costimulatory molecules such as CD80, CD86 or CD40, thus lacking immunogenicity. Furthermore, they are able to suppress T- and B-lymphocyte activation and proliferation and may also affect dendritic cell maturation. Based on these properties, MSC are being used in regenerative medicine and also for the treatment of autoimmune diseases and graft-versus-host disease. On the other...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mesenchymal stem cells; T-lymphocytes; B-lymphocytes; Dendritic cells; Graft-versus-host disease; Allogeneic transplanta.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000500002
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Isolation and culture of umbilical vein mesenchymal stem cells BJMBR
Covas,D.T.; Siufi,J.L.C.; Silva,A.R.L.; Orellana,M.D..
Bone marrow contains a population of stem cells that can support hematopoiesis and can differentiate into different cell lines including adipocytes, osteocytes, chondrocytes, myocytes, astrocytes, and tenocytes. These cells have been denoted mesenchymal stem cells. In the present study we isolated a cell population derived from the endothelium and subendothelium of the umbilical cord vein which possesses morphological, immunophenotypical and cell differentiation characteristics similar to those of mesenchymal stem cells isolated from bone marrow. The cells were isolated from three umbilical cords after treatment of the umbilical vein lumen with collagenase. The cell population isolated consisted of adherent cells with fibroblastoid morphology which, when...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Mesenchymal stem cells; Umbilical vein; Adipocytes; Osteocytes; Cell differentiation.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000900006
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Mesenchymal Stem Cell treatment for autoimmune diseases: a critical review Biol. Res.
Figueroa,Fernando E; Carrión,Flavio; Villanueva,Sandra; Khoury,Maroun.
Mesenchymal stem cells (MSCs) are now known to display not only stem cell multipotency, but also robust antiinflammatory and regenerative properties. After widespread in-vitro and in-vivo preclinical testing, autologous and allogeneic MSCs have been applied in a range of immune mediated conditions, including graft versus host disease, Crohn's disease, multiple sclerosis, refractory systemic lupus erythematosus and systemic sclerosis. Current data suggests that MSCs may not only replace diseased tissues, but also exert several trophic, regenerative and antiinflammatory effects. While the clinical outcome in case reports and phase I-II trials seems occasionally striking, these limited results point to the need to perform controlled multicenter trials. Future...
Tipo: Journal article Palavras-chave: Mesenchymal stem cells; Autoimmune diseases; Cellular therapy.
Ano: 2012 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602012000300008
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Mesenchymal stem cells for the treatment of neurodegenerative and psychiatric disorders Anais da ABC (AABC)
COLPO,GABRIELA D.; ASCOLI,BRUNA M.; WOLLENHAUPT-AGUIAR,BIANCA; PFAFFENSELLER,BIANCA; SILVA,EMILY G.; CIRNE-LIMA,ELIZABETH O.; QUEVEDO,JOÃO; KAPCZINSKI,FLÁVIO; ROSA,ADRIANE R..
Mesenchymal stem cells (MSCs) are multipotent progenitor cells that have the capacity to differentiate into all lineages of mesodermal origin, e.g., cartilage, bone, and adipocytes. MSCs have been identified at different stages of development, including adulthood, and in different tissues, such as bone marrow, adipose tissue and umbilical cord. Recent studies have shown that MSCs have the ability to migrate to injured sites. In this regard, an important characteristic of MSCs is their immunomodulatory and anti-inflammatory effects. For instance, there is evidence that MSCs can regulate the immune system by inhibiting proliferation of T and B cells. Clinical interest in the use of MSCs has increased considerably over the past few years, especially because...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mesenchymal stem cells; Treatment; Neurodegenerative disease; Psychiatric disorders.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652015000301435
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Mesenchymal stem cells from patients with chronic myeloid leukemia do not express BCR-ABL and have absence of chimerism after allogeneic bone marrow transplant BJMBR
Carrara,R.C.V.; Orellana,M.D.; Fontes,A.M.; Palma,P.V.B.; Kashima,S.; Mendes,M.R.; Coutinho,M.A.; Voltarelli,J.C.; Covas,D.T..
Bone marrow is a heterogeneous cell population which includes hematopoietic and mesenchymal progenitor cells. Dysregulated hematopoiesis occurs in chronic myelogenous leukemia (CML), being caused at least in part by abnormalities in the hematopoietic progenitors. However, the role of mesenchymal stem cells (MSCs) in CML has not been well characterized. The objectives of the present study were to observe the biological characteristics of MSCs from CML patients and to determine if MSCs originate in part from donors in CML patients after bone marrow transplantation (BMT). We analyzed MSCs from 5 untreated patients and from 3 CML patients after sex-mismatched allogeneic BMT. Flow cytometry analysis revealed the typical MSC phenotype and in vitro assays showed...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mesenchymal stem cells; Chimerism; Allogeneic hematopoietic stem cell transplantation; Chronic myeloid leukemia; Bcr-abl gene.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2007000100008
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MSC transplantation: a promising therapeutic strategy to manage the onset and progression of diabetic nephropathy Biol. Res.
Ezquer,Marcelo E; Ezquer,Fernando E; Arango-Rodríguez,Martha L; Conget,Paulette A.
Currently, one of the main threats to public health is diabetes mellitus. Its most detrimental complication is diabetic nephropathy (DN), a clinical syndrome associated with kidney damage and an increased risk of cardiovascular disease. Irrespective of the type of diabetes, DN follows a well-known temporal course. The earliest detectable signs are microalbuminuria and histopathological changes including extracellular matrix deposition, glomerular basement membrane thickening, glomerular and mesangial expansion. Later on macroalbuminuria appears, followed by a progressive decline in glomerular filtration rate and the loss of glomerular podocytes, tubulointerstitial fibrosis, glomerulosclerosis and arteriolar hyalinosis. Tight glycemic and hypertension...
Tipo: Journal article Palavras-chave: Regenerative medicine; Diabetes mellitus; Diabetic nephropathy; Multipotent mesenchymal stromal cells; Mesenchymal stem cells.
Ano: 2012 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602012000300010
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Overlapping of mononuclear cells derived from bone marrow in rats' intervertebral discs: an in vitro study Ciência Rural
Fontes,Erica Batista; Borges,Andréa Pacheco Batista; Pippi,Ney Luis; Rosa,Maurício; Sprada,Arícia; Togni,Monique.
In this study, bone marrow mononuclear cells (BM-MNC) derived from rats were used in order to promote intervertebral disc regeneration. These cells were isolated after centrifugation in a Ficoll-Paque™ PLUS density gradient and then placed in plastic dishes to proliferate during a period of 14 days. The BM-MNCs were previously labeled with the fluorescent membrane marker Chloromethyl-benzamidodialkylcarbocyanine (CM-DIL), and thereafter were implanted in rats' intervertebral discs explants as an in vitro experimental model. Daily analyses of the cells under a fluorescence microscope revealed morphological changes, which assumed a thin and elongated shape similar to cells that originally form the annulus fibroses. Histopathological analysis demonstrated the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mesenchymal stem cells; Intervertebral disc; In vitro; Rat.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782010000400024
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Poly(trimethylene carbonate-co-L-lactide) electrospun scaffolds for use as vascular grafts BJMBR
Braghirolli,D.I.; Caberlon,B.; Gamba,D.; Petry,JFTC.; Dias,M.L.; Pranke,P..
Currently, there is great clinical need for suitable synthetic grafts that can be used in vascular diseases. Synthetic grafts have been successfully used in medium and large arteries, however, their use in small diameter vessels is limited and presents a high failure rate. In this context, the aim of this study was to develop tissue engineering scaffolds, using poly(trimethylene carbonate-co-L-lactide) (PTMCLLA), for application as small diameter vascular grafts. For this, copolymers with varying trimethylene carbonate/lactide ratios – 20/80, 30/70, and 40/60 – were submitted to electrospinning and the resulting scaffolds were evaluated in terms of their physicochemical and biological properties. The scaffolds produced with PTMCLLA 20/80, 30/70, and 40/60...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Poly(trimethylene carbonate-co-L-lactide); Vascular scaffolds; Mesenchymal stem cells; Endothelial progenitor cells; Smooth muscle cells.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2019000800611
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Rat dental pulp stem cells: isolation and phenotypic characterization method aiming bone tissue bioengineering BABT
Bertassoli,Bruno Machado; Costa,Emanuela Silva; Sousa,Cristiane Aparecida; Albergaria,Juliano Douglas Silva; Maltos,Kátia L. Melo; Goes,Alfredo Miranda; Matins,Thais Maria da Mata; Silva,Gerluza Aparecida Borges; Jorge,Erika Cristina.
ABSTRACT: Dental pulp stem cells (DPSC) have been showing a considerable potential for regenerative medicine. Pulps were collected from lower incisors (n=2) through direct access of the tooth pulp chamber. The isolated cells were cultured in alfa-MEM 10% FBS, in standard culture conditions. At the third passage, DPSC were characterized by flow cytometry (MHCI, CD54, CD73, CD90, CD45, CD11 and CD34); RT-PCR for Nanog gene; and their differentiation capacity in osteogenic, adipogenic and chondrogenic cell lines. Isolated cells exhibited adhesion capacity to plastic; fusiform morphology, and 80% confluence reached in approximately 3 days. These cells have also revealed positive expression for CD54, CD73 and CD90 markers; and negative expression for CD11, CD34...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mesenchymal stem cells; Dental pulp; Murine; Phenotypic characterization; Cellular differentiation.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100335
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The epigenetic modifiers 5-aza-2'-deoxycytidine and trichostatin A influence adipocyte differentiation in human mesenchymal stem cells BJMBR
Zych,J.; Stimamiglio,M.A.; Senegaglia,A.C.; Brofman,P.R.S.; Dallagiovanna,B.; Goldenberg,S.; Correa,A..
Epigenetic mechanisms such as DNA methylation and histone modification are important in stem cell differentiation. Methylation is principally associated with transcriptional repression, and histone acetylation is correlated with an active chromatin state. We determined the effects of these epigenetic mechanisms on adipocyte differentiation in mesenchymal stem cells (MSCs) derived from bone marrow (BM-MSCs) and adipose tissue (ADSCs) using the chromatin-modifying agents trichostatin A (TSA), a histone deacetylase inhibitor, and 5-aza-2′-deoxycytidine (5azadC), a demethylating agent. Subconfluent MSC cultures were treated with 5, 50, or 500 nM TSA or with 1, 10, or 100 µM 5azadC for 2 days before the initiation of adipogenesis. The differentiation was...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mesenchymal stem cells; Trichostatin A; 5-Aza-2′-deoxycytidine; Adipocyte differentiation; Epigenetics.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2013000500405
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The TLR7 agonist Imiquimod promote the immunogenicity of msenchymal stem cells Biol. Res.
Zhang,Li; Liu,Dan; Pu,Dan; Wang,Yanwen; Li,Li; He,Yanqi; Li,Yalun; Li,Lei; Li,Weimin.
BACKGROUND: Mesenchymal stem cells (MSCs) are considered the best candidate in stem cells therapy due to their multipotent differentiation ability, low expression of co-stimulatory molecules (CD80, CD86, CD34 and HLA-II) and immunosuppression effects on in vivo immune responses. MSCs were now widely used in clinical trials but received no encourage results. The major problem was the fate of engrafted MSCs in vivo could not be defined. Some studies indicated that MSCs could induce immune response and result in the damage and rejection of MSCs. As toll like receptors (TLRs) are important in inducing of immune responses, in this study we study the role of TLR7 in mediating the immune status of MSCs isolated from umbilical cord. RESULTS: Our results indicated...
Tipo: Journal article Palavras-chave: Mesenchymal stem cells; Umbilical cord; Toll like receptor 7; Immunogenicity; Pro-inflammatory molecules.
Ano: 2015 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100006
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Using mesenchymal stem cells as a therapy for bone regeneration and repairing Biol. Res.
Shao,Jin; Zhang,Weiwei; Yang,Tieyi.
Bone is a unique tissue which could regenerate completely after injury rather than heal itself with a scar. Compared with other tissues the difference is that, during bone repairing and regeneration, after the inflammatory phase the mesenchymal stem cells (MSCs) are recruited to the injury site and differentiate into either chondroblasts or osteoblasts precursors, leading to bone repairing and regeneration. Besides these two precursors, the MSCs can also differentiate into adipocyte precursors, skeletal muscle precursors and some other mesodermal cells. With this multiline-age potentiality, the MSCs are probably used to cure bone injury and other woundings in the near future. Here we will introduce the recent developments in understanding the mechanism of...
Tipo: Journal article Palavras-chave: Mesenchymal stem cells; Bone regeneration; Bone repairing; Tissue engineering.
Ano: 2015 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100062
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Variation of Hydroxyapatite Content in Soft Gelatin Affects Mesenchymal Stem Cell Differentiation BABT
Kantawong,Fahsai; Tanum,Junijira; Wattanutchariya,Wassanai; Sooksaen,Pat.
Gelatin is a common material used in tissue engineering and hydroxyapatite (HA) has a composition and structure similar to natural bone mineral. HA is also used to increase the adhesion ability of scaffolds. The physical and mechanical properties of gelatin, together with the chemical properties of HA, can affect cell differentiation. The main purpose of this study is to investigate the gene expression of human mesenchymal stem cells (HMSCs) upon culturing on gelatin composite with HA. Low amounts of HA were introduced into the gelatin in order to modulate properties of gelatin. Three types of hydrogel were fabricated by glutaraldehyde crosslinking before lyophilization to produce the porous 3D structure: (1) pure gelatin, (2) 0.5 mg/ml HA in gelatin, and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Gelatin; Hydroxyapatite; Mesenchymal stem cells; Gene Expression.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100411
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