(OP 145) Human Cultured Epithelial Allografts: an Adapted Approach Introducing More Efficient Production Schemes and Resulting in Extra Patient Safety

• Published in: Tissue engineering. Part A Vol. 14; no. 5; p. 741
• Main Authors: De Corte, P, Verween, G, Verbeken, G, Rose, T, Jennes, S, Pirnay, J-P
• Format: Journal Article
• Language: English
• Published: 01.05.2008
• ISSN: 1937-3341; 1937-335X

Introduction: Since 1987 keratinocytes are grown and used in our burn unit (> 800 patients were treated). During those two decades, our culture system has evolved considerably with an emphasis on safety assurance for the acceptor patients, increased quality and overall performance, and cost-reduction. Methods: The major improvements consist in the introduction of neonatal foreskin keratinocytes (NFK), a feeder layer- and animal component free culture system, and a Quality Management System (QMS). Results: In contrast to the irregular and unpredictable growth rates and end stages of normal adult human epidermal keratinocytes (HEKa), NFKs show a very regular and consistent growth, comparable to that of established cell lines, which lasts for up to 50 population doublings (theoretical expansion of a 1 cm super(2) biopsy up to 100 km super(2) of cultured epithelium, compared to 50 m super(2) for adult keratinocytes). This tremendous proliferation potential allows the use of more strict and predictable subculturing schemes. Today, all culturing and banking procedures are performed taking into account the cGMP guidelines and ISO 9001:2000 QMS requirements. Recently, our lab tested of a new totally defined and animal free culture medium. This medium supported a regular and extended growth (up to 25 population doublings). The use of a totally defined and animal free medium is avoiding possible transmittance of potential animal viral agents. The support for the cells is given by a coating of recombinant human type-1 collagen instead of the traditional bovine collagen. We also tested (safe) membranes as a support for human epithelial grafts.