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  • Growth and stratification of epithelial cells in minimal culture conditions.

Growth and stratification of epithelial cells in minimal culture conditions.

Methods in molecular biology (Clifton, N.J.) (2009-11-13)
Federica Riva, Andrea Casasco, Marco Casasco, Alberto Calligaro, Antonia Icaro Cornaglia
ABSTRACT

Biological risk management is required in modern tissue engineering. Particular attention should be paid to the culture medium and the scaffold used. In this perspective, it is important to define minimal culture conditions which allow proper growth and differentiation of epithelial cells in vitro. We propose a simple experimental system which permits the generation of three-dimensional epidermal constructs using a collagen layer as a scaffold mimicking the entire dermal tissue and without the need of any feeder layer. Although showing significant differences compared to natural epidermis, these epidermal constructs appear useful to study keratinocyte differentiation and epidermis histogenesis.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Collagen from calf skin, Bornstein and Traub Type I, solid, BioReagent, suitable for cell culture
Sigma-Aldrich
apo-Transferrin bovine, BioReagent, suitable for cell culture, ≥98%
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Epoxy embedding medium, hardener MNA, ≥95.0%
Sigma-Aldrich
Pepsin from porcine gastric mucosa, powder, ≥400 units/mg protein
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Monoclonal Anti-Laminin antibody produced in mouse, clone LAM-89, ascites fluid
Sigma-Aldrich
Goat serum