Many studies are being conducted to define the role of growth factors in cutaneous physiology in order to add cytokines in a timely fashion for optimal tissue engineering of skin. This study is aimed at developing a multistep approach for the production of bioengineered skin substitutes, taking into account the effects of various growth factors according to the culture time. The use of a serum-supplemented medium throughout the whole culture period of skin substitutes was compared to the sequential use of specific additives at defined culture steps. Histological analysis revealed that serum was necessary for keratinocyte proliferation and migration on dermal substitutes during the first 2 d after their seeding. However, the serum-free medium presented some advantages when supplemented with different additives at specific culture steps. Interestingly, ascorbic acid added to the dermal substitutes before and after keratinocyte seeding maintained their cuboïdal morphology in the basal epidermal layer. In the absence of serum, collagen matrix degradation slowed down, and a better multilayered epidermal organization was obtained, notably with retinoic acid. Stratum corneum formation was also enhanced by fatty acids. Thus, sequential addition of exogenous factors to the medium used to produce skin substitutes can improve their structural features and functional properties in vitro.
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1 February 2000
MULTISTEP PRODUCTION OF BIOENGINEERED SKIN SUBSTITUTES: SEQUENTIAL MODULATION OF CULTURE CONDITIONS
FRANÇOIS A. AUGER,
ROXANE POULIOT,
NATHALIE TREMBLAY,
RINA GUIGNARD,
PATRICIA NOËL,
JULIANNA JUHASZ,
LUCIE GERMAIN,
FRANCINE GOULET
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In Vitro Cellular & Developmental Biology - Animal
Vol. 36 • No. 2
February 2000
Vol. 36 • No. 2
February 2000
bioengineering
culture
Skin substitute