In Australia, approximately 2,000 children/year are hospitalized due to burn injuries and the after-effects can last a lifetime. While new burn wound therapies are tested on animal models, this process is expensive and ethically questionable. A 3D, composite, in vitro skin model would enable rapid testing of new potential treatments. Dermal replacement matrices (DRM) are used surgically to replace skin in deep burn injuries. These DRM are readily available and could be utilised as a human “in vitro” skin model. Primary human keratinocyte and fibroblast skin cells were grown on four matrices (Biobrane®, Biodegradable-Temporising Matrix – BTM, Integra® and Matriderm®) to develop tissue similar to native human skin. Different ratios of the primary cells were seeded onto the matrices to determine the cell numbers required for optimal growth. Fluorescent staining confirmed the presence of living and dead cells within the matrices. Histological dyes were used to observe morphology and immunohistochemistry was used to characterize and localize the cells. After the model was confirmed to be similar to native human skin, it was used as an “in vitro human skin” burn model and the staining techniques described above were used to characterise the cell damage. Integra® and Matriderm® had the most suitable cell adherence and proliferation. Biobrane®, BTM and Matriderm® withstood heat up to 90° without deformation. The most appropriate DRM for an in vitro skin burn model was Matriderm®, as it supported superior cell growth and withstood the heat of burning.