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Generation of a three-dimensional full thickness skin equivalent and automated wounding

: Rossi, Angela; Appelt-Menzel, Antje; Kurdyn, Szymon; Walles, Heike; Groeber, Florian


Journal of visualized experiments : JoVE. Online resource (2015), Nr.96, Art. e52576
ISSN: 1940-087X
Fraunhofer IGB ()
tissue engineering; 3D in vitro models; test systems; alternative to animal testing; full thickness; skin equivalent; skin injury; wound model; automation; wounding device

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models reflect the human wound situation better than animal models. Although two-dimensional models are widely used to investigate processes such as cellular migration and proliferation, models that are more complex are required to gain a deeper knowledge about wound healing. Besides a suitable model system, the generation of precise and reproducible wounds is crucial to ensure comparable results between different test runs. In this study, the generation of a three-dimensional full thickness skin equivalent to study wound healing is shown. The dermal part of the models is comprised of human dermal fibroblast embedded in a rat-tail collagen type I hydrogel. Following the inoculation with human epidermal keratinocytes and consequent culture at the air-liquid interface, a multilayered epidermis is formed on top of the models. To study the wound healing process, we additionally developed an automated wounding device, which generates standardized wounds in a sterile atmosphere.