Publication: Human keratinocytes cultured without a feeder layer undergo progressive loss of differentiation markers
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Date
1999
Authors
Prignano, F. ; Domenici Lombardo, L. ; Gerlini, G. ; Pimpinelli, N. ; Romagnoli, P.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
Culture of keratinocytes in conventional
medium without a mesenchyme-derived feeder layer
leads to poor growth and impaired differentiation;
however, the exact pathway and degree of differentiation
achieved in such conditions is unclear. We have cultured
normal human keratinocytes in Rheinwald and Green's
medium, on plastic without a feeder layer, in order to
investigate the degree of differentiation that they achieve
in these conditions. Intermediate filament proteins,
tonofibrils and desmosomes were assumed as markers of
differentiation and their expression was analyzed by
immunohistochemistry and electron micros~opy. Before
reaching confluence, keratinocytes expressed keratin
molecules, as well as vimentin, and formed tonofibrils
and desmosomes. The expression of these markers was
progressively reduced until confluence and was totally
lost thereafter, while cultures could be propagated for at
least six passages. On the contrary, reseeding on a feeder
layer after the first passage led to rapid cell death. It
could be concluded that signals from a feeder layer are
relevant to support continuous synthesis of intermediate
filaments proteins and formation of tonofibils and
desmosomes, and that the derangement of the cytoskeleton
in these conditions leads to altered, not simply
defective, response to delayed stimulation by a feeder
layer.
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