Optimization and characterization of an engineered human skin equivalent
Optimization and characterization of an engineered human skin equivalent
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DOI：
10.1089/ten.2005.11.723
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发表时间：
2005-05-01
影响因子：
--
通讯作者：
Damour, O
中科院分区：
文献类型：
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作者：
Black, AF;Bouez, C;Damour, O
皮肤等效物（SEs）的设计可以满足基础研究和应用研究的需要。组织工程se的成功应用要求重组组织具有正确的组织结构和功能。大量的实验证据现在支持这样一种观点，即除了细胞间相互作用外，间充质组织对上皮细胞形态发生的诱导作用至少部分是由细胞外基质成分介导的。采用成纤维细胞和角质形成细胞共培养模型，研究渐进式血清减少对表皮分化、真皮和真皮-表皮连接质量以及细胞外基质蛋白表达的影响。将细胞依次添加到由胶原蛋白、糖胺聚糖和壳聚糖组成的真皮底物中。本研究的主要目的是优化该模型用于药物毒理学试验。对照皮肤等效物在整个生产过程中用含10%血清的培养基培养。在气液界面处将血清含量降至1%和0%，并与对照皮肤当量进行比较。首先，我们证明了在气液界面处血清剥夺可改善角化细胞终末分化。其次，我们发现，在没有血清的情况下，SE的特异性特征得以维持，包括表皮和真皮的超微结构，真皮主要细胞外基质成分（人I、III和V型胶原、纤维连接蛋白、弹性蛋白和原纤维蛋白1）的表达，以及真皮-表皮连接点（层粘连蛋白、人IV型胶原、α(6)整合素）的表达。此外，我们的结果表明，使用角化细胞和成纤维细胞的共培养模型具有生物学有用组织所需的形态和功能特性。
Skin equivalents (SEs) have been designed to meet both basic and applied research needs. The successful application of tissue-engineered SEs requires that the reconstituted tissues be endowed with the correct organization and function. A large body of experimental evidence now supports the notion that the inducing effects of mesenchymal tissue on epithelial cell morphogenesis are mediated, at least in part, by extracellular matrix components in addition to cell-cell interactions. A coculture model including both fibroblasts and keratinocytes was used to study the effects of progressive serum reduction on epidermal differentiation, quality of dermal and dermal-epidermal junctions, and expression of extracellular matrix proteins. The cells were successively added to a dermal substrate composed of collagen, glycosaminoglycans, and chitosan. The main aim of this study was to optimize this model for pharmacotoxicological trials. Control skin equivalents were cultured with medium containing 10% serum throughout the production process. Serum content was reduced to 1 and 0% at the air-liquid interface and compared with control skin equivalents. First, we demonstrated that serum deprivation at the air-liquid interface improves keratinocyte terminal differentiation. Second, we showed that, in the absence of serum, the specific characteristics of the SE are maintained, including epidermal and dermal ultrastructure, the expression of major dermal extracellular matrix components ( human collagen types I, III, and V, fibronectin, elastin, and fibrillin 1), and the dermal-epidermal junction ( laminin, human type IV collagen, alpha(6) integrin). Furthermore, our results indicate that coculture models using keratinocytes and fibroblasts have both morphological and functional properties required for biologically useful tissues.