Reconstructed human skin models are a valuable tool for drug discovery, disease modeling, and basic research. In the past decades, major progress has been made in this field leading to the development of full-thickness skin models (FTSms) better representative of the native human skin by including the cellular cross talk between the dermal and epidermal layers. However, current available FTSms still present important limitations since they are only suitable for short-term studies, include nonhuman extracellular matrix (ECM) components and have a weak skin barrier function compared with in vivo human skin. In this study, a fibroblast-derived matrix was combined with the use of an inert polystyrene scaffold for the development of a fully human dermis capable of supporting a differentiated epidermis. To produce a pigmented FTSm, a coculture with keratinocytes, melanocytes, and fibroblasts was established. The structure and functionality of the developed FTSms were studied for short- and long-term cultivation using histological and immunofluorescence staining. The integrity of the skin barrier was evaluated using transepithelial electrical resistance (TEER) measurements. It was possible to obtain a mature dermis capable of supporting an epidermis without keratinocyte infiltration in only 6 days. ECM components (collagen IV and fibrin) were secreted by the fibroblasts and accumulated in the scaffold structure, recreating the microenvironment of the native human dermis. Moreover, the use of a scaffold resulted in a structure with mechanical stability due to its noncontracting nature. The coculture of primary human keratinocytes resulted in a terminally differentiated skin equivalent that could maintain its architecture and homeostasis up to 50 days. Melanocytes were correctly integrated within the epidermal basal layer and made it possible to reproduce constitutive pigmentation. TEER levels increased during culture time, reaching values of 1.1 ± 0.2 kΩ.cm² for the FTSm, indicative of a functional skin barrier.

中文翻译：

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用于长期研究的基于成纤维细胞衍生矩阵的有色全层人体皮肤模型

重建的人体皮肤模型是药物发现、疾病建模和基础研究的宝贵工具。在过去的几十年中，该领域取得了重大进展，通过包括真皮层和表皮层之间的细胞串扰，开发了更能代表天然人类皮肤的全层皮肤模型 (FTSms)。然而，目前可用的 FTSms 仍然存在重要的局限性，因为它们仅适用于短期研究，包括非人类细胞外基质 (ECM) 成分并且与体内相比具有较弱的皮肤屏障功能人的皮肤。在这项研究中，成纤维细胞衍生基质与惰性聚苯乙烯支架的使用相结合，以开发能够支持分化表皮的全人真皮。为了产生有色素的 FTSm，建立了与角质形成细胞、黑色素细胞和成纤维细胞的共培养。使用组织学和免疫荧光染色研究了开发的 FTSms 的结构和功能，用于短期和长期培养。使用跨上皮电阻 (TEER) 测量来评估皮肤屏障的完整性。仅在 6 天内就有可能获得能够支撑表皮而没有角质形成细胞浸润的成熟真皮。ECM 成分（胶原蛋白 IV 和纤维蛋白）由成纤维细胞分泌并积累在支架结构中，重建天然人类真皮的微环境。此外，由于其非收缩性质，支架的使用导致具有机械稳定性的结构。原代人类角质形成细胞的共培养产生了终末分化的皮肤等效物，可以维持其结构和稳态长达 50 天。黑色素细胞被正确地整合到表皮基底层中，并使其有可能再现组成性色素沉着。TEER 水平在培养期间增加，达到 1.1 ± 0.2 kΩ.cm 的值 原代人类角质形成细胞的共培养产生了终末分化的皮肤等效物，可以维持其结构和稳态长达 50 天。黑色素细胞被正确地整合到表皮基底层中，并使其有可能再现组成性色素沉着。TEER 水平在培养期间增加，达到 1.1 ± 0.2 kΩ.cm 的值 原代人类角质形成细胞的共培养产生了终末分化的皮肤等效物，可以维持其结构和稳态长达 50 天。黑色素细胞被正确地整合到表皮基底层中，并使其有可能再现组成性色素沉着。TEER 水平在培养期间增加，达到 1.1 ± 0.2 kΩ.cm 的值FTSm 为²，表示功能性皮肤屏障。