In vitro co-culture of human skin keratinocytes and fibroblasts on a biocompatible and biodegradable scaffold.
نویسندگان
چکیده
BACKGROUND Extensive full-thickness burns require replacement of both epidermis and dermis. In designing skin replacements, the goal has been to re-create this model and make a product which has both essential components. METHODS In the present study, we developed procedures for establishing confluent, stratified layers of cultured human keratinocytes on the surface of modified collagen-chitosan scaffold that contains fibroblasts. The culture methods for propagation of keratinocytes and fibroblasts isolated from human neonatal foreskin were developed. The growth and proliferation of normal human keratinocytes were evaluated in serum-free (keratinocyte growth medium) and our modified medium. Characterization of human keratinocytes was determined by using pan-keratin and anti-involucrin monoclonal antibodies. For fabrication of relevant biodegradable and biocompatible collagen-chitosan porous scaffold with improved biostability, modified method of freeze-gelation was used. In generating organotypic co-cultures, epidermal keratinocytes were plated onto the upper surface of scaffold containing embedded fibroblasts. RESULTS The results showed that the growth of isolated human skin fibroblasts and keratinocytes in our modified medium was more than that in the serum-free medium. The different evaluations of collagen-chitosan scaffold showed that it is relevant to growth of cells (fibroblast and keratinocyte) and has a good flexibility in manipulation of the living skin equivalents. CONCLUSION These findings indicate that the integration of collagen-chitosan scaffold with co-cultured keratinocyte and fibroblast in vitro provides a potential source of living skin for grafting in vivo.
منابع مشابه
In vitro Co-Culture of Human Skin Keratinocytes and Fibroblasts on a Biocompatible and Biodegradable Scaffold
Background: Extensive full-thickness burns require replacement of both epidermis and dermis. In designing skin replacements, the goal has been to re-create this model and make a product which has both essential components. Methods: In the present study, we developed procedures for establishing confluent, stratified layers of cultured human keratinocytes on the surface of modified collagen-chito...
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Abstract Background: Considering the ineffective conventional therapeutic methods for treating different types of wounds, and because of reports of successful use of skin substitutes prepared from co-cultured keratinocytes and fibroblasts on the scaffold of collagen, this study was performed to evaluate the skin substitute on experimentally induced wounds in rats. Materials and methods: Thi...
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ورودعنوان ژورنال:
- Iranian biomedical journal
دوره 13 3 شماره
صفحات -
تاریخ انتشار 2009