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Enhancement of keratinocyte performance in the production of tissue‐engineered skin using a low‐calcium medium
Author(s) -
Her Catherine A.,
Harrison Caroline A.,
Thornton Daniel J. A.,
MacNeil Sheila
Publication year - 2007
Publication title -
wound repair and regeneration
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.847
H-Index - 109
eISSN - 1524-475X
pISSN - 1067-1927
DOI - 10.1111/j.1524-475x.2007.00275.x
Subject(s) - keratinocyte , egta , epidermis (zoology) , calcium , dermis , wound healing , tissue engineering , tissue culture , chemistry , organ culture , microbiology and biotechnology , regeneration (biology) , in vitro , biomedical engineering , medicine , pathology , biochemistry , immunology , anatomy , biology , organic chemistry
ABSTRACT The success of laboratory‐expanded autologous keratinocytes for the treatment of severe burn injuries is often compromised by their lack of dermal remnants and failure to establish a secure dermo‐epidermal junction on the wound bed. We have developed a tissue‐engineered skin substitute for in vivo use, based on a sterilized donor human dermis seeded with autologous keratinocytes and fibroblasts. However, culture rates are currently too slow for clinical use in acute burns. Our aim in this study was to increase the rate of production of tissue‐engineered skin. Two approaches were explored: one using a commercial low‐calcium media and the other supplementing well‐established media for keratinocyte culture with the calcium‐chelating agent ethylene glutamine tetra‐acetic acid (EGTA). Using commercial low‐calcium media for both the initial cell culture and subsequent culture of tissue‐engineered skin did not produce tissue suitable for clinical use. However, it was possible to enhance the initial proliferation of keratinocytes and to increase their horizontal migration in tissue‐engineered skin by supplementing established culture medium with 0.04 mM EGTA without sacrificing epidermal attachment and differentiation. Enhancement of keratinocyte migration with EGTA was also maximal in the absence of fibroblasts or basement membrane.