Human keratinocytes cultured on collagen matrix used as an experimental burn model.

Abstract

Background: In experimental models in vivo, it is difficult to characterize the effect of thermal burns on epidermal keratinocytes. Since the response to thermal injury involves several systemic mechanisms, especially because of the stimulus to coagulation and inflammatory cascades, it becomes hard to evaluate the specific effect of thermal burns on keratinocytes. The aim of this study is to propose the use of human keratinocytes cultured on collagen matrix as an in vitro experimental burn model.

Methods: Human keratinocytes derived from neonatal foreskins were isolated and cultured following standard methods. All experiments used the same keratinocyte lineage and were carried out in triplicate. Initially, gels of collagen and Matrigel were prepared. For each gel, 2 x 10(6) keratinocytes were seeded and cultured to form stratified epithelia. Following, burn wounds were induced at 170 degrees C.

Results: Keratinocytes were cultured on collagen-coated Millicell membranes. Stratified epithelia were formed and burned on the seventh day after the cultures were raised to the air-liquid interface. The burn procedure is reproducible and can be easily executed.

Conclusion: The proposed model can be used to study the effects of induced burn wounds on keratinocytes in a specific way.