Generation of human epidermal constructs on a collagen layer alone

Federica Riva, Andrea Casasco, Emanuele Nespoli, Antonia Icaro Cornaglia, Marco Casasco, Angela Faga, Silvia Scevola, Giuliano Mazzini, Alberto Calligaro

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Because engineered tissues are designed for clinical applications in humans, a major problem is the contamination of cocultures and tissues by allogenic molecules used to grow stem cells in vitro. The protocols that are commonly applied to generate epidermal equivalents in vitro require the use of irradiated murine fibroblasts as a feeder layer for keratinocytes. In this study, we report a simple procedure for growing human keratinocytes, isolated from adult skin, to generate an epidermal construct on a collagen layer alone. In this model, no human or murine feeder layers were used to amplify cell growth, and isolated keratinocytes were seeded directly at high cell density on the collagen-coated flasks or coverslips in an epithelial growth medium containing low calcium concentration. Morphological, immunochemical, and cytokinetic features of epithelial colonies grown on the collagen layer were typical of keratinocytes and were comparable with those reported for keratinocytes grown on a feeder layer. The stratification of keratinocytes generated 3-dimensional synthetic constructs displaying a tissue architecture comparable with that of natural epidermis. Epithelial cells expressed specific markers of keratinocyte terminal differentiation, including involucrin and filaggrin. Nevertheless, the number of cell layers was lower than in natural skin, and electron microscopical analysis revealed that the overall organization of these layers was poor compared with natural epidermis, including the formation of junctional complexes, basement membrane, and keratinization. The lack of epithelial-mesenchymal interactions that occur during skin histogenesis may account for such an incomplete maturation of epidermal constructs.

Original languageEnglish
Pages (from-to)2769-2779
Number of pages11
JournalTissue Engineering
Volume13
Issue number11
DOIs
Publication statusPublished - Nov 1 2007

Fingerprint

Keratinocytes
Collagen
Feeder Cells
Skin
Tissue
Epidermis
Cell Count
Cell growth
Fibroblasts
Growth
Coculture Techniques
Stem cells
Basement Membrane
Calcium
Contamination
Stem Cells
Epithelial Cells
Cells
Electrons
Molecules

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Biotechnology

Cite this

Riva, F., Casasco, A., Nespoli, E., Cornaglia, A. I., Casasco, M., Faga, A., ... Calligaro, A. (2007). Generation of human epidermal constructs on a collagen layer alone. Tissue Engineering, 13(11), 2769-2779. https://doi.org/10.1089/ten.2006.0329

Generation of human epidermal constructs on a collagen layer alone. / Riva, Federica; Casasco, Andrea; Nespoli, Emanuele; Cornaglia, Antonia Icaro; Casasco, Marco; Faga, Angela; Scevola, Silvia; Mazzini, Giuliano; Calligaro, Alberto.

In: Tissue Engineering, Vol. 13, No. 11, 01.11.2007, p. 2769-2779.

Research output: Contribution to journalArticle

Riva, F, Casasco, A, Nespoli, E, Cornaglia, AI, Casasco, M, Faga, A, Scevola, S, Mazzini, G & Calligaro, A 2007, 'Generation of human epidermal constructs on a collagen layer alone', Tissue Engineering, vol. 13, no. 11, pp. 2769-2779. https://doi.org/10.1089/ten.2006.0329
Riva F, Casasco A, Nespoli E, Cornaglia AI, Casasco M, Faga A et al. Generation of human epidermal constructs on a collagen layer alone. Tissue Engineering. 2007 Nov 1;13(11):2769-2779. https://doi.org/10.1089/ten.2006.0329
Riva, Federica ; Casasco, Andrea ; Nespoli, Emanuele ; Cornaglia, Antonia Icaro ; Casasco, Marco ; Faga, Angela ; Scevola, Silvia ; Mazzini, Giuliano ; Calligaro, Alberto. / Generation of human epidermal constructs on a collagen layer alone. In: Tissue Engineering. 2007 ; Vol. 13, No. 11. pp. 2769-2779.
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