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The effect of serum withdrawal on the protein profile of quiescent human dermal fibroblasts in primary cell culture.Boraldi Federica,
Annovi Giulia,
Paolinelli Devincenzi Chiara,
Tiozzo Roberta,
Quaglino Daniela, 2008, izvirni znanstveni članek
Opis: The effect of serum deprivation on proliferating cells is well known, in contrast its role on primary
cell cultures, at confluence, has not been deeply investigated. Therefore, in order to explore
the response of quiescent cells to serum deprivation, ubiquitous mesenchymal cells, as normal
human dermal fibroblasts, were grown, for 48 h after confluence, in the presence or absence of
10% FBS. Fibroblast behaviour (i.e. cell morphology, cell viability, ROS production and elastin
synthesis) was evaluated morphologically and biochemically. Moreover, the protein profile was
investigated by 2-DE and differentially expressed proteins were identified by MS. Serum withdrawal
caused cell shrinkage but did not significantly modify the total cell number. ROS production,
as evaluated by the dihydroethidium (DH2) probe, was increased after serum deprivation,
whereas elastin synthesis, measured by a colorimetric method, was markedly reduced in the
absence of serum. By proteome analysis, 41 proteins appeared to significantly change their
expression, the great majority of protein changes were related to the cytoskeleton, the stress response
and the glycolytic pathway. Data indicate that human dermal fibroblasts in primary cell
culture can adapt themselves to environmental changes, without significantly altering cell viability,
at least after a few days of treatment, even though serum withdrawal represents a stress
condition capable to increase ROS production, to influence cell metabolism and to interfere with
cell behaviour, favouring the expression of several age-related features.
Ključne besede: Dermal fibroblasts / Primary cell culture / ROS production / Serum withdrawal
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