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首页> 外文期刊>Biomaterials >Multi-layered culture of human skin fibroblasts and keratinocytes through three-dimensional freeform fabrication.
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Multi-layered culture of human skin fibroblasts and keratinocytes through three-dimensional freeform fabrication.

机译：通过三维自由形式制作的人皮肤成纤维细胞和角质形成细胞的多层培养。

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摘要

We present a method to create multi-layered engineered tissue composites consisting of human skin fibroblasts and keratinocytes which mimic skin layers. Three-dimensional (3D) freeform fabrication (FF) technique, based on direct cell dispensing, was implemented using a robotic platform that prints collagen hydrogel precursor, fibroblasts and keratinocytes. A printed layer of cell-containing collagen was crosslinked by coating the layer with nebulized aqueous sodium bicarbonate. The process was repeated in layer-by-layer fashion on a planar tissue culture dish, resulting in two distinct cell layers of inner fibroblasts and outer keratinocytes. In order to demonstrate the ability to print and culture multi-layered cell-hydrogel composites on a non-planar surface for potential applications including skin wound repair, the technique was tested on a poly(dimethylsiloxane) (PDMS) mold with 3D surface contours as a target substrate. Highly viable proliferation of each cell layer was observed on both planar and non-planar surfaces. Our results suggest that organotypic skin tissue culture is feasible using on-demand cell printing technique with future potential application in creating skin grafts tailored for wound shape or artificial tissue assay for disease modeling and drug testing.
机译：我们提出一种方法来创建由人类皮肤成纤维细胞和模拟皮肤层的角质形成细胞组成的多层工程组织复合材料。使用直接打印胶原蛋白水凝胶前体，成纤维细胞和角质形成细胞的机器人平台，实现了基于直接细胞分配的三维（3D）自由形式制造（FF）技术。通过用雾化的碳酸氢钠水溶液涂布该层，使含有细胞的胶原蛋白的印刷层交联。在平面组织培养皿上以逐层方式重复该过程，从而产生内部成纤维细胞和外部角质形成细胞两个不同的细胞层。为了展示在非平面表面上印刷和培养多层细胞-水凝胶复合材料以进行潜在的应用（包括皮肤伤口修复）的能力，该技术在具有3D表面轮廓的聚二甲基硅氧烷（PDMS）模具上进行了测试目标衬底。在平面和非平面表面上均观察到每个细胞层的高度增殖。我们的研究结果表明，使用按需细胞印刷技术进行器官型皮肤组织培养是可行的，未来可能在创建针对伤口形状定制的皮肤移植物或用于疾病建模和药物测试的人工组织测定中潜在应用。

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