Kolundzic Nikola, Khurana Preeti, Crumrine Debra, Celli Anna, Mauro Theodora M, Ilic Dusko
Department of Women & Children's Health, School of Life Course & Population Sciences, Faculty of Life Sciences & Medicine, King's College London, London, United Kingdom.
Assisted Conception Unit (ACU), Guy's and St Thomas' NHS Foundation Trust, London, United Kingdom.
JID Innov. 2021 Dec 7;2(2):100083. doi: 10.1016/j.xjidi.2021.100083. eCollection 2022 Mar.
Epidermal basement membrane, a tightly packed network of extracellular matrix (ECM) components, is a source of physical, chemical, and biological factors required for the structural and functional homeostasis of the epidermis. Variations within the ECM create distinct environments, which can affect the property of cells in the basal layer of the epidermis and subsequently affect keratinocyte differentiation and stratification. Very little attention has been paid to mimicking basement membrane in organotypic cultures. In this study, using parameters outlined in a consensus on the quality standard of organotypic models suitable for dermatological research, we have evaluated three basement membrane substitutes. We compared fibronectin with three complex three-dimensional matrices: Matrigel, decellularized dermal fibroblast‒produced and ‒assembled ECM, and a dry human amniotic membrane. Our results suggest that Matrigel is not a suitable substrate for human epidermal equivalent culture, whereas the two other complex three-dimensional substitutes, decellularized dermal fibroblast‒produced and ‒assembled ECM and dry human amniotic membrane, were superior to single layer fibronectin coating. Human epidermal equivalents cultured on either decellularized dermal fibroblast‒produced and ‒assembled ECM or on dry human amniotic membrane generated hemidesmosomes, whereas those on fibronectin did not. In addition, human epidermal equivalent cultured on decellularized dermal fibroblast‒produced and ‒assembled ECM and on dry human amniotic membrane can be maintained in culture 4 days longer than human epidermal equivalent cultured on fibronectin without compromising the barrier function.
表皮基底膜是细胞外基质（ECM）成分紧密排列的网络，是表皮结构和功能稳态所需的物理、化学和生物因子的来源。ECM内的变化创造了不同的环境，这会影响表皮基底层细胞的特性，进而影响角质形成细胞的分化和分层。在器官型培养中模拟基底膜的研究很少受到关注。在本研究中，我们使用适用于皮肤病学研究的器官型模型质量标准共识中概述的参数，评估了三种基底膜替代物。我们将纤连蛋白与三种复杂的三维基质进行了比较：基质胶、脱细胞真皮成纤维细胞产生并组装的ECM以及干燥的人羊膜。我们的结果表明，基质胶不是人表皮替代物培养的合适底物，而另外两种复杂的三维替代物，即脱细胞真皮成纤维细胞产生并组装的ECM和干燥的人羊膜，优于单层纤连蛋白涂层。在脱细胞真皮成纤维细胞产生并组装的ECM或干燥的人羊膜上培养的人表皮替代物产生了半桥粒，而在纤连蛋白上培养的则没有。此外，在脱细胞真皮成纤维细胞产生并组装的ECM和干燥的人羊膜上培养的人表皮替代物在培养中比在纤连蛋白上培养的人表皮替代物可多维持4天，且不损害屏障功能。
