ǂThe ǂeffect of serum withdrawal on the protein profile of quiescent human dermal fibroblasts in primary cell culture

Federica Boraldi (Avtor), Giulia Annovi (Avtor), Chiara Paolinelli DeVincenzi (Avtor), Roberta Tiozzo (Avtor), Daniela Quaglino (Avtor)

Povzetek

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;

Podatki

Jezik:	Angleški jezik
Leto izida:	2008
Tipologija:	1.01 - Izvirni znanstveni članek
Organizacija:	UNG - Univerza v Novi Gorici
UDK:	54
COBISS:	5425659
ISSN:	1615-9861
Št. ogledov:	2658
Št. prenosov:	0
Ocena:	0 (0 glasov)
Metapodatki:

Ostali podatki

URN:	URN:SI:UNG
Vrsta dela (COBISS):	Delo ni kategorizirano
Strani:	str. 66-82
Letnik:	ǂVol. ǂ8
Zvezek:	ǂno. ǂ1
Čas izdaje:	2008
DOI:	10.1002/pmic.200700833
ID:	11192840