P23 Critical evaluation of animal-component-free media in human keratinocytes and human fibroblasts cell culture

Abstract Introduction and aims Many classic three-dimensional (3D) skin models contain animal-derived materials e.g. bovine/rat collagen I. In this pioneering project, animal-derived materials were replaced with animal-component-free materials to evaluate the cellular interactions so that an animal-component-free full-thickness 3D skin model can be created. The first step to creating an animal-component-free 3D skin model is to change the culture medium of the cells. There are defined animal-component-free media available; however, they can be expensive. This project evaluates the most beneficial cost-effective media. The aim of this project was to evaluate whether animal-component-free media affects human primary skin cells compared with cells grown in animal-derived media in a two-dimensional cell culture. Methods Normal human dermal fibroblasts (NHDFs) and normal human epidermal keratinocytes (NHEKs) were grown in animal-component-free media and animal-derived media. The cells were treated with both commercial defined media or in-house animal-component-free-media containing various different supplements including growth hormones and cytokines and left to grow for 7 days. On days 3, 5 and 7, the morphology, cell growth and viability were analysed. The morphology was imaged using a Leica DMi8 microscope. From this, average cell size was calculated using Image J. The cells were counted with trypan blue in an automated cell counter for cell growth and viability analysis. Expression studies and an enzyme-linked immunosorbent assay were done with cellular material. An Anova test was performed for statistical significance. Results The results show that as the duration of culture increased, the cells grown in animal-component-free media had an increased growth rate (in-house media and commercial defined media) compared with animal-derived media. There was a change in the morphology of primary skin cells grown in different animal-component-free media. Conclusions This project analyses the effects of animal-component-free media compared with animal-derived media on NHEKs and NHDFs, examining multiple factors including cellular growth and gene expression.