New skin-equivalent model from de-epithelialized amnion membrane
New skin-equivalent model from de-epithelialized amnion membrane
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DOI：
10.1007/s00441-006-0208-2
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发表时间：
2006-10-01
影响因子：
3.6
通讯作者：
Hashimoto, Koji
中科院分区：
文献类型：
--
作者：
Yang, Lujun;Shirakata, Yuji;Hashimoto, Koji
在构建人类活体皮肤等效物（LSE）时，已有基底膜（BM）成分的存在改善了表皮和基底膜的形态发生。去上皮化羊膜（AM）保留了关键的BM成分。因此，我们研究了AM对构建LSE的有用性。去上皮AM覆盖在包埋成纤维细胞的I型胶原凝胶上。然后将正常人角质形成细胞（NHKs）播种到AM的上皮侧以构建AM- lse。通过在成纤维细胞填充的I型胶原凝胶上播种NHKs构建常规LSE。当角质形成细胞达到融合时，将LSE举至气液界面，培养3周。在不同的时间采集样品，并进行形态学、免疫组织化学和超微结构的研究。在AM-LSE中，表皮分层更好，基底细胞更致密，极化，柱状，分化和增殖标志物的表达与正常人皮肤更相似。AM-LSE组BM更连续，半粒酶更发达。AM-LSE的表皮生长速度明显快于未添加AM的LSE。当移植到裸鼠身上时，两种lse效果良好；然而，AM-LSE移植物表皮、基底膜和半底粒的形态发生较好。在体外和体内实验中，AM-LSE具有更好的表皮形态和更发达的基底膜，这表明AM-LSE在临床应用上优于不加AM的LSE。
The presence of pre-existing basement membrane (BM) components improves the morphogenesis of epidermis and BM in constructing a human living skin-equivalent (LSE). De-epithelialized amniotic membrane (AM) retains key BM components. We have therefore investigated the usefulness of AM for constructing LSE. De-epithelialized AM was overlaid on type I collagen gel embedded with fibroblasts. Normal human keratinocytes (NHKs) were then seeded onto the epithelial side of the AM to construct an AM-LSE. A conventional LSE was constructed by seeding NHKs on a fibroblast-populated type I collagen gel. When the keratinocytes reached confluence, the LSE was lifted to the air-liquid interface and cultured for up to 3 weeks. Samples were harvested at various times and investigated morphologically, immunohistochemically, and ultrastructurally. In AM-LSE, the epidermis was better stratified, with more compact, polarized, columnar basal cells, and the expression of differentiation and proliferation markers was more similar to that of normal human skin than was that of LSE without AM. A more continuous BM and better-developed hemidesmosomes were found in AM-LSE. The epidermis of AM-LSE outgrew much faster than that of LSE without AM. When transplanted onto nude mice, both LSEs took well; however, the AM-LSE graft showed better morphogenesis of the epidermis, BM, and hemidesmosomes. The better epidermal morphology and better-developed BM in AM-LSE in vitro and in vivo indicates its superiority over LSE without AM for clinical applications.