Proteins present within the cell layer and those released in the cell medium from in vitro cultured normal human dermal fibroblasts were separated and characterized in terms of their isoelectric point and molecular weight, by two-dimensional (2-D) gel electrophoresis. All spots in the synthetic gel were firstly analyzed by the Melanie 3 software and compared with those of breast cancer cells, colorectal epithelial cells, HL60, lymphoma cells, and platelets, already available on-line. From the identification of 144 spots from both the cell layer and the medium, we were able to recognize 89 different proteins, since a certain number of spots represented different isoforms of the same molecule. Identifications were performed by matching with on-line 2-D databases, and by matrix assisted laser-desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS), in order to confirm the identification by matching, or to identify new proteins. The procedure we used allows (i) to design a highly reproducible reference map of the proteome of adult human normal fibroblasts in culture, (ii) to evaluate protein species produced in the cell layer as well as those released in the culture medium, and (iii) to compare data from gel matching with those obtained by MS. This work represents an essential step for a better knowledge of mesenchymal cells, given the widespread use of this cell type in both clinical and experimental investigations.

Normal human dermal fibroblasts: Proteomic analysis of cell layer and culture medium / Boraldi, Federica; L., Bini; S., Liberatori; A., Armini; V., Pallini; Tiozzo, Roberta; Ronchetti, Ivonne; Quaglino, Daniela. - In: ELECTROPHORESIS. - ISSN 0173-0835. - STAMPA. - 24:7-8(2003), pp. 1292-1310. [10.1002/elps.200390166]

Normal human dermal fibroblasts: Proteomic analysis of cell layer and culture medium

BORALDI, Federica;TIOZZO, Roberta;RONCHETTI, Ivonne;QUAGLINO, Daniela
2003

Abstract

Proteins present within the cell layer and those released in the cell medium from in vitro cultured normal human dermal fibroblasts were separated and characterized in terms of their isoelectric point and molecular weight, by two-dimensional (2-D) gel electrophoresis. All spots in the synthetic gel were firstly analyzed by the Melanie 3 software and compared with those of breast cancer cells, colorectal epithelial cells, HL60, lymphoma cells, and platelets, already available on-line. From the identification of 144 spots from both the cell layer and the medium, we were able to recognize 89 different proteins, since a certain number of spots represented different isoforms of the same molecule. Identifications were performed by matching with on-line 2-D databases, and by matrix assisted laser-desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS), in order to confirm the identification by matching, or to identify new proteins. The procedure we used allows (i) to design a highly reproducible reference map of the proteome of adult human normal fibroblasts in culture, (ii) to evaluate protein species produced in the cell layer as well as those released in the culture medium, and (iii) to compare data from gel matching with those obtained by MS. This work represents an essential step for a better knowledge of mesenchymal cells, given the widespread use of this cell type in both clinical and experimental investigations.
2003
24
7-8
1292
1310
Normal human dermal fibroblasts: Proteomic analysis of cell layer and culture medium / Boraldi, Federica; L., Bini; S., Liberatori; A., Armini; V., Pallini; Tiozzo, Roberta; Ronchetti, Ivonne; Quaglino, Daniela. - In: ELECTROPHORESIS. - ISSN 0173-0835. - STAMPA. - 24:7-8(2003), pp. 1292-1310. [10.1002/elps.200390166]
Boraldi, Federica; L., Bini; S., Liberatori; A., Armini; V., Pallini; Tiozzo, Roberta; Ronchetti, Ivonne; Quaglino, Daniela
File in questo prodotto:
Non ci sono file associati a questo prodotto.
Pubblicazioni consigliate

Licenza Creative Commons
I metadati presenti in IRIS UNIMORE sono rilasciati con licenza Creative Commons CC0 1.0 Universal, mentre i file delle pubblicazioni sono rilasciati con licenza Attribuzione 4.0 Internazionale (CC BY 4.0), salvo diversa indicazione.
In caso di violazione di copyright, contattare Supporto Iris

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11380/305593
Citazioni
  • ???jsp.display-item.citation.pmc??? 4
  • Scopus 19
  • ???jsp.display-item.citation.isi??? 19
social impact