title: Plasticity of protein expression during culture of fetal skin cells.
abstract: In order to gain insight into the biology of fetal skin during culture, cellular proteins were studied during four culture passages (P00, P01, P04 as well as P10) using high-resolution two-dimensional (2-D) gel electrophoresis and mass spectrometry (MS). Bioinformatic analyses were focused on a region of each gel corresponding to pI between 4 and 8 and M(r) from 8000 to 35 000. In this area, 373 +/- 42 spots were detected (N = 18). Twenty-six spots presented an integrated intensity that increased in the higher passages, whereas five spots showed a progressively lower intensity in subsequent passaging. MS analysis was performed on spots that were unambiguously identified on preparative 2-D gels. Among the 26 spots showing an increased size between P00 and P10, 9 were identified, and corresponded to 3 proteins: (i) peptidyl-prolyl cis-trans isomerase A (P05092; cyclophilin A or cyclosporin A-binding protein), (ii) triosephosphate isomerase (P00938), and (iii) enoyl-CoA hydratase (P30084). Among these nine identified spots, three were absent at P00, but were present at P10. They corresponded to isoforms of peptidyl-prolyl cis-trans isomerase and triosephosphate isomerase, respectively. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses of the acidic isoforms of triosephosphate isomerase showed modifications of cysteine residues to cysteic acid. All these isoforms were clearly present in the skin cells of a 4-year-old child, as well as in skin cells from a 80-year-old man, at P00. These observations probably reflect either an oxidative stress related to cell culture, or, alternatively, maturation, differentiation and the aging of the cells.
authors.affiliation: Service Régional Vaudois de Transfusion Sanguine, Lausanne, Switzerland.
authors.forename: Françoise
authors.initials: F
authors.lastname: Vuadens
authors.forename: David
authors.initials: D
authors.lastname: Crettaz
authors.forename: Corinne
authors.initials: C
authors.lastname: Scelatta
authors.forename: Catherine
authors.initials: C
authors.lastname: Servis
authors.forename: Manfredo
authors.initials: M
authors.lastname: Quadroni
authors.forename: Willy-Vincent
authors.initials: WV
authors.lastname: Bienvenut
authors.forename: Philippe
authors.initials: P
authors.lastname: Schneider
authors.forename: Patrick
authors.initials: P
authors.lastname: Hohlfeld
authors.forename: Lee A
authors.initials: LA
authors.lastname: Applegate
authors.forename: Jean-Daniel
authors.initials: JD
authors.lastname: Tissot
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D002478:Cells, Cultured; D002675:Child, Preschool; D021984:Cyclophilin A / Q000032:analysis; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004746:Enoyl-CoA Hydratase / Q000032:analysis; D005333:Fetus; D015870:Gene Expression; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D014176:Protein Biosynthesis; D020033:Protein Isoforms / Q000032:analysis; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D012867:Skin / Q000166:cytology* / Q000196:embryology; D023822:Tissue Engineering; D014305:Triose-Phosphate Isomerase / Q000032:analysis
chemical_list: D020033:Protein Isoforms; D011506:Proteins; D004746:Enoyl-CoA Hydratase; D021984:Cyclophilin A; D014305:Triose-Phosphate Isomerase