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Muscle-derived stem cells in tissue engineering: defining cell properties suitable for construct design.

The terms construct or tissue equivalent refer to neotissue produced by tissue engineering techniques. The elements forming the construct are scaffolds on which cells are "recreated" to form an engineered-tissue sensitive to certain cell signals. The ability of the cells to expand and differentiate on the scaffold is determined by properties such as fixation, adhesion, proliferation and migration. Among the cell types that seem to be most promising for designing constructs are tissue-residing, or adult, stem cells, which show two main features: a capacity to differentiate into many cell lineages and the power of self-renewal. These features make them good candidates for cell replacement therapies. Here, we report the identification, isolation and culture of muscle stem cells aimed at establishing the ideal culture in terms of defining when the cultured cell population would show optimal characteristics for transfer to the scaffold to obtain a particular construct. Stem cells harvested from the dorsal muscle of white New Zealand rabbits were cultured in vitro and characterized 5 to 14 days after the start of culture. Fibroblasts obtained from the same experimental animal served as controls. The stem cells were examined by light and scanning electron microscopy. For stem cell identification, we used the antibodies anti-m-cadherin, anti-CD34 and anti-Myf-5. The markers of muscle differentiation used were: anti-vimentin, anti-alpha-actin, anti-desmin and anti-myosin. The expression profiles of the different markers of muscle differentiation and TGFbeta1 in the cell cultures were confirmed by Western blotting. Proliferation rates were determined by monitoring tritiated thymidine incorporation. The thymidine incorporation rate was substantially higher for the population of undifferentiated cells than for control fibroblasts obtained from the same animal. During the first five days of culture, most cells were negative for all the markers examined, with the exception of m-cadherin, CD34 and Myf-5, although discrete signs of vimentin expression started to emerge. After 14 days of culture, the adult stem cells showed vimentin (94.2%) and desmin (33.8%) expression yet scarce labeling for myosin (16.2%) and alpha-actin (8.3%). Control fibroblasts showed intense labeling for vimentin (99.3%) and alpha-actin (62.2%), while less than 2% of the population expressed myosin (0.9%) and desmin (1.6%). After two weeks of culture, muscle-derived stem cells show good proliferative and adhesion properties as they initiate differentiation. These conditions seem ideal for obtaining the desired construct.
J Buján, G Pascual, C Corrales, V Gómez-Gil, M Rodríguez, J M Bellón

1474 related Products with: Muscle-derived stem cells in tissue engineering: defining cell properties suitable for construct design.

10 ug1 mg100ul96 assays1x10e7 cells1.00 flask

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Detection of atherosclerotic plaque with two monoclonal antibodies. 2P1A2 monoclonal antibody is specific for smooth muscle cells in atherosclerotic plaque.

We investigated two monoclonal antibodies (MAbs) which recognize rabbit atherosclerotic tissues. In particular, one antibody, 2P1A2, is specific for rabbit aortic smooth muscle cells (RASMCs). We used RASMCs in primary culture to produce and screen MAbs directed towards the cell surface. The specificity of the described antibodies was tested on a battery of tissue cryosections of different origin (rabbit, rat and human) by immunological staining. 2P1A2 shows an exclusive immunolabeling for SMCs present inside rabbit atherosclerotic plaque. This MAb shows inside the fibrous plaque a staining similar to two other SMC-specific antibodies (anti-desmin and anti-alpha-actin). In an early stage of atherosclerosis, close to the internal elastic lamina, underlying a fibrous plaque, 2P1A2 detects some SMCs; in contrast, anti-desmin and anti-alpha-actin fail to stain such SMCs. This antibody may be therefore considered as directed specifically against SMCs in an activated state. The other antibody which we describe, 1PC1, stains a pericellular antigen expressed by cultured SMCs and shows a specificity for smooth muscle tissues. 1PC1 MAb strongly stains the fibrous plaque of atherosclerotic rabbit aorta and the recognized epitope is present inside the aortic media. These two antibodies may be useful in the recognition of vascular SMCs during the atherosclerotic process.
J M Lamaziere, A Desmouliere, M Pascal, J Larrue

2313 related Products with: Detection of atherosclerotic plaque with two monoclonal antibodies. 2P1A2 monoclonal antibody is specific for smooth muscle cells in atherosclerotic plaque.

0.2 mg100 ug25 µg25 µg100 TESTS1 ml0.25 mg100ug Lyophilized1 mg200 ug200 ug100 ul

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Immunocytochemical study of intermediate filaments in cultured human trabecular cells.

In order to study which classes of intermediate filaments (IF) comprise the IF of human trabecular cells, we undertook immunocytochemical investigations on cultured human trabecular cells using anti-keratin, anti-vimentin, anti-desmin and anti-glial fibrillar acidic protein (GFAP) rabbit sera. Immunostaining with the keratin antibody and the GFAP antibody only produced nonspecific staining. Immunostaining with the vimentin antibody and the desmin antibody produced intracytoplasmic filamentous staining. Pretreatment of the cells with colcemid to induce the perinuclear concentration of IF resulted in the demonstration of a characteristic perinuclear immunofluorescence, confirming the existence of vimentin and desmin. Some cells showed particularly strong positivity to desmin. The desmin antibody used in this study only faintly labeled the IF of cultured human skin fibroblasts, known to contain vimentin but not desmin. These results suggest that the IF of human trabecular cells are composed of vimentin and desmin, and that human trabecular cells possess muscle cell-like functions.
Y Iwamoto, M Tamura

2528 related Products with: Immunocytochemical study of intermediate filaments in cultured human trabecular cells.

1.00 flask1 mg10 ug100 extractions1.00 flask1.00 flask1.00 flask1 mg96 wells (1 kit)100 ul100 μg2ug

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