Eves Paula C, Bullett Nial A, Haddow David, Beck Alison J, Layton Christopher, Way Louise, Shard Alex G, Gawkrodger David J, Mac Neil Sheila
Department of Engineering Materials, The Kroto Research Institute, North Campus, University of Sheffield, Sheffield, UK.
J Invest Dermatol. 2008 Jun;128(6):1554-64. doi: 10.1038/sj.jid.5701214. Epub 2008 Jan 17.
Obtaining pigmentary function in autologous skin grafts is a current challenge for burn surgeons as is developing reliable robust grafting strategies for patients with vitiligo and piebaldism. In this paper, we present the development of a simple methodology for delivering cultured keratinocytes and melanocytes to the patient that is of low risk for the patient but also user friendly for the surgeon. In this study, we examined the ability of keratinocytes and melanocytes to transfer from potential cell carriers under different media conditions to an in vitro human wound bed model. The number of melanocytes transferred, their location within the neoepidermis, and their ability to pigment were evaluated as preclinical end points. Two inert substrates (polyvinyl chloride and silicone sheets) and three candidate plasma-polymerized coatings with controlled surface chemistry deposited on these substrates were explored. Two media for expansion of cells, Greens, currently used clinically (but which contains fetal calf serum), and a serum-free alternative, M2 (melanocyte medium), were explored. Reproducible transfer of physiologically relevant numbers of melanocytes capable of pigmentation from the coculture of melanocytes and keratinocytes was obtained using either Greens medium or M2 medium, and a silicone carrier pretreated with 20% carboxylic acid deposited by plasma polymerization.
在自体皮肤移植中实现色素功能,对于烧伤外科医生来说是一项当前的挑战,就如同为白癜风和斑驳病患者开发可靠且稳健的移植策略一样。在本文中,我们展示了一种简单方法的开发,该方法用于将培养的角质形成细胞和黑素细胞递送至患者,对患者风险低且对外科医生来说操作简便。在本研究中,我们检测了在不同培养基条件下,角质形成细胞和黑素细胞从潜在细胞载体转移至体外人伤口床模型的能力。转移的黑素细胞数量、它们在新表皮内的位置以及它们的色素沉着能力被评估为临床前终点。探索了两种惰性底物(聚氯乙烯和硅胶片)以及三种沉积在这些底物上具有可控表面化学性质的候选等离子体聚合涂层。探索了两种用于细胞扩增的培养基,目前临床上使用的格林培养基(但含有胎牛血清)以及一种无血清替代品M2(黑素细胞培养基)。使用格林培养基或M2培养基,以及经等离子体聚合沉积20%羧酸预处理的硅胶载体,可实现从黑素细胞和角质形成细胞共培养物中可重复转移具有色素沉着能力的生理相关数量的黑素细胞。