Publication: Muscle-derived stem cells in tissue engineering:
defining cell properties suitable for construct design
Authors
Buján, J. ; Pascual, G. ; Corrales, C. ; Gómez-Gil, V. ; Rodríguez, M. ; Bellón, J.M.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
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 enginnered-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: antivimentin,
anti-a-actin, anti-desmin and anti-myosin. The
expression profiles of the different markers of muscle
differentiation and TGFß1 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 a-actin (8.3%).
Control fibroblasts showed intense labeling for vimentin
(99.3%) and a-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.
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