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FUTURE WORK • The use of this simplified cell culture procedure will translate into a greater capacity for studies of bioengineered human skin. • Increasing knowledge in this field will enable discovery of next-generation bioengineered skin to assist the warfighter Disclaimer: Neither the authors or their family members have a financial interest in any commercial product, service, or organization providing financial support for this research, nor do they endorse any products or entities mentioned. The opinions and assertions expressed herein are those of the authors and do not necessarily reflect the official policy or position of the Uniformed Services University or the Department of Defense. REFERENCES 1. Thangapazham RL, Darling TN, Meyerle J. Alteration of skin properties with autologous dermal fibroblasts. Int J Mol Sci. 2014;15(5):8407-27. 2. Thangapazham RL, Klover P, Wang JA, Zheng Y, Devine A, Li S, Sperling L, Cotsarelis G, Darling TN. Dissociated human dermal papilla cells induce hair follicle neogenesis in grafted dermal-epidermal composites. J Invest Dermatol. 2014;134:538-540. A Simplified Culture Protocol that Preserves Hair Follicle Neogenesis in Grafted Dermal-Epidermal Constructs Jahnabi Roy1,2, Donald Aduba, Jr.1,2, Shaowei Li 1,2, Jordan Betz3,4 , Ji-an Wang 1,2, Vincent B. Ho 3,5 Thomas Darling1 1Department of Dermatology, Uniformed Services University, Bethesda, MD 2 Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD 3 4D Bio 3 Center for Biotechnology, Uniformed Services University, Bethesda, MD 4 The Geneva Foundation, Tacoma, WA 5 Department of Radiology and Radiological Sciences, Uniformed Services University Bethesda, MD SPECIFIC AIMS  To investigate effects of CnT-Prime Full Thickness 3D Airlift Medium (PRIME AIRLIFT) compared to Epi and Cori media on: 1) In vitro proliferation of dermal papilla cells and keratinocytes, and expression of alkaline phosphatase, a marker of trichogenicity. 2) DEC graft size a, RITM0028638, ? Large, full-thickness skin wounds heal with scars that have impaired skin function and appearance, caused in part by absence of hair follicles [1]. ? Hair follicles produce hair and sebum and are a storehouse of epidermal stem cells. ? Current methods to promote healing of large, full-thickness skin wounds do not promote regeneration of hair follicles: ?Autologous split-thickness skin grafts ? Skin substitutes, including bioengineered dermalepidermal composites (DECs) made of fibroblasts embedded in a collagen matrix layered with keratinocytes. ?We achieved human hair follicle neogenesis in DECs composed of human neonatal foreskin keratinocytes and human dermal papilla cells [2]. ? In vitro cultivation involved the sequential use of two types of media with complicated formulations: epidermalization (Epi) and cornification (Cori) media ? In this study, we examined different types of commercially available media in order to simplify in vitro cultivation and in an attempt to enhance hair follicle formation.