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| Dielectrophoresis: a review of applications for stem cell research.
May 22, 2010 at 8:32 AM
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| | Dielectrophoresis: a review of applications for stem cell research. J Biomed Biotechnol. 2010;2010:182581 Authors: Pethig R, Menachery A, Pells S, De Sousa P Dielectrophoresis can discriminate distinct cellular identities in heterogeneous populations, and monitor cell state changes associated with activation and clonal expansion, apoptosis, and necrosis, without the need for biochemical labels. Demonstrated capabilities include the enrichment of haematopoetic stem cells from bone marrow and peripheral blood, and adult stem cells from adipose tissue. Recent research suggests that this technique can predict the ultimate fate of neural stem cells after differentiation before the appearance of specific cell-surface proteins. This review summarises the properties of cells that contribute to their dielectrophoretic behaviour, and their relevance to stem cell research and translational applications. PMID: 20490279 [PubMed - in process] | | |
| The interactions between rASCs, rhBMP-2 and beta-TCP play an important role in bone tissue engineering.
May 22, 2010 at 8:32 AM
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| | The interactions between rASCs, rhBMP-2 and beta-TCP play an important role in bone tissue engineering. Tissue Eng Part A. 2010 May 20; Authors: E LL, Xu L, Wu X, Wang D, Lv Y, Wang J, Liu H ABSTRACT Cells, scaffolds, and growth factors are the three main factors for creating a stem cell-based tissue engineered construct, but the interactions between three factors are not very clear. We hereby explored the interactions between rat adipose-derived stromal cells (rASCs), bone morphogenetic protein-2 (rhBMP-2) and beta-tricalcium phosphate (ss-TCP) to provide evidence for their application in bone tissue engineering by evaluating the protein adsorption of ss-TCP, the cell attachment, alkaline phosphatase (ALP) activity/protein, osteocalcin content, mineral formation, calcium content, phosphonium content, cell vitality, gene expression and implantation in the backs of severe combined immunodeficient (SCID) mice of rhBMP-2 pre-inducing rASCs seeded onto ss-TCP. The attachment, proliferation and osteogenic properties of rASCs were supported by ss-TCP using scanning electron microscope (SEM); Compared with rASCs cultured on the culture plate, rASCs cultlured! on ss-TCP had significantly higher ALP activity/protein, osteocalcin content and mineral formation, the values for rASCs cultured on ss-TCP with rhBMP-2 increased most significantly; The rhBMP-2 significantly increased calcium content, phosphonium content and ALP, I type collagen, osteocalcin mRNAs levels of rASCs cultured on ss-TCP; After 8 and 12 weeks of implantation, each group displayed increased bone formation over the 12-week time period. The value for rASCs/ss-TCP construct was significantly higher than that for ss-TCP group. But the maximal and robust bone formation was presented in rASCs/ss-TCP with rhBMP-2. The results implied that stem cells existed in adult rat adipose tissue. ss-TCP could adsorb rhBMP-2 from the media and had osteoinductivity when alone implanted in the back of SCID mice. ss-TCP was also sufficient to trigger the differentiation of rASCs toward an osteoblastic phenotype without the addition of osteogenic factor. The rhBMP-2 could better suffi! ciently induce osteogenic differentiation of rASCs seeded onto! ss-TCP. The rASCs and rhBMP-2 could promote the dissolution of ss-TCP to provide Ca2+, PO3-4 needed by bone formation. The interactions between three factors could provide an optimizing microenvironment for osteogenic differentiation of rASCs, this might be essential for sufficient and timely bone formation in vivo. This study may provide insight into the clinical repair of bone defect with ASCs+ss-TCP+rhBMP-2 construct. PMID: 20486786 [PubMed - as supplied by publisher] | | |
| Non expanded mesenchymal stem cells for regenerative medicine: yield in stromal vascular fraction from adipose tissues.
May 22, 2010 at 8:32 AM
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| | Non expanded mesenchymal stem cells for regenerative medicine: yield in stromal vascular fraction from adipose tissues. Tissue Eng Part C Methods. 2010 May 20; Authors: Faustini M, Bucco M, Chlapanidas T, Lucconi G, Marazzi M, Tosca M, Gaetani P, Klinger M, Villani S, Ferretti VV, Vigo D, Torre ML The adipose-derived stromal vascular fraction (SVF) represents a rich source of mesenchymal cells, potentially able to differentiate into adipocytes, chondrocytes, osteoblasts, myocytes, cardiomyocytes, hepatocytes, and neuronal, epithelial and endothelial cells. These cells are ideal candidates for use in regenerative medicine, tissue engineering, including gene therapyand cell replacement cancer therapies. In this work, aiming to the optimization of the adipose stem cell-based therapy, the effect of the collection site, surgical procedure and tissue processing techniques on SVF yield was evaluated in terms of cell recovery and live cells, taking into account the effect of gender, age and BMI. Adipose tissue samples were recovered from 125 informed subjects (37 males and 88 females, mean age: 51.31 y, range: 15-87 y), and digested in different condition with collagenase. A multivariate linear model put in evidence that the gender of the donor should be taken into! account, as well as the collection site and the tissue digestion conditions; the collection technique, the age and the body mass index of donor seem not to influence the cell yield. PMID: 20486782 [PubMed - as supplied by publisher] | | |
| Human Bone-Marrow And Adipose Tissue Mesenchymal Stem Cells: A User's Guide.
May 22, 2010 at 8:32 AM
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| | Human Bone-Marrow And Adipose Tissue Mesenchymal Stem Cells: A User's Guide. Stem Cells Dev. 2010 May 20; Authors: Mosna F, Sensebé L, Krampera M Mesenchymal Stem Cells (MSCs) are adult stem cells which hold great promise in the field of Regenerative Medicine. They can be isolated from almost any tissue of the body and display, after expansion, very similar properties and minor differences, probably due to their microenvironment of origin. Expansion in vitro can be obtained in cytokine-free, serum-enriched media, as well as in serum-free, basic Fibroblast Growth Factor-enriched media. A detailed immunophenotypic analysis is required to test the purity of the preparation, but no unique distinguishing marker has been described as yet. Functional assays, i.e. differentiation studies in vitro, are needed to prove multilineage differentiation of expanded cells, and demonstration of pluripotency is necessary to identify most immature precursors. MSCs show powerful immunomodulative properties towards most of the cells of the immune system: this strengthens the theoretical rationale for their use also in an allogen! eic setting across the MHC immunological barriers. Systemic intravenous injection, as well as local use have been tried: following systemic injection, MSCs show a high degree of chemotaxis based on pro-inflammatory cytokines, and localize at inflamed and neoplastic tissues; local regeneration has been improved using synthetic, as well as organic scaffolds. On the other hand, inadequate heterotopic in vivo differentiation and neoplastic transformation are potential risks of this form of cell therapy, even if evidence of this sort have been collected only from studies in mice, and generally after prolonged in vitro expansion. This review tries to provide a detailed technical overview of the methods used for human bone marrow-derived and adipose tissue-derived MSCs isolation, in vitro expansion and characterization for tissue repair. We chose to use BM-MSCs as a model to describe techniques that have been used for MSCs isolation and expansion from very different sources, and A! T-MSCs as an example of a reliable and increasingly common alt! ernative source. PMID: 20486777 [PubMed - as supplied by publisher] | | | | 
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