Abstract Tissue engineering is emerging as a modern medicine fascination towards the establishment of human tissue banks; yet, these approaches typically involve cultures of only one type of cell and, therefore, do not recapitulate the native tissue physiology in toto. Co-culture models, comprised of different cell types, can potentially create the next level of complexity. However, conventional approaches involving multiple cell types and cell culture inserts do have limitations. To this end, here we demonstrate a novel cell culture insert that allows the use of any custom-made scaffold, free-flow of fluids/gases, dual cell seeding on either sides of the insert, easy stacking of multiple inserts and resizing it to any multi-well plate format as well as culture dishes. To prove the concept, electrospun silk fibroin scaffold was clipped onto the insert and was used for co-culturing of keratinocytes and fibroblast cells. The results indicated a successful fabrication of spatially organized skin tissue constructs having epidermal and dermal equivalent histology. Cell-laden inserts were stacked and used for simulated transportation studies. However, the conditions need further fine-tuning. All together, the results indicated that the novel cell culture insert with silk fibroin scaffold could be used as a facile, versatile and scalable approach to fabricate and transport 3D co-cultured tissue constructs in vitro, including but not limited to skin. The resultant tissue constructs can be explored for therapeutic applications, for instance as artificial skin substitute in wound healing, and for toxicological applications, for instance as reconstructed skin tissue model in skin irritation testing.

中文翻译：

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使用具有夹式支架的双细胞接种兼容细胞培养插入物的共培养组织构建体的生物制造，用于潜在的再生医学和毒理学筛选应用

摘要 组织工程正在成为现代医学对建立人体组织库的迷恋；然而，这些方法通常只涉及一种类型细胞的培养，因此不能完整地概括天然组织的生理学。由不同细胞类型组成的共培养模型可能会产生更高的复杂性。然而，涉及多种细胞类型和细胞培养插入物的传统方法确实有局限性。为此，我们在这里展示了一种新型细胞培养插入物，它允许使用任何定制的支架、流体/气体的自由流动、插入物两侧的双细胞接种、多个插入物的轻松堆叠并调整其大小任何多孔板形式以及培养皿。为了证明这个概念，电纺丝素蛋白支架夹在插入物上，用于角化细胞和成纤维细胞的共培养。结果表明具有表皮和真皮等效组织学的空间组织的皮肤组织构建体的成功制造。将载有细胞的插入物堆叠起来并用于模拟运输研究。但是，条件需要进一步微调。总之，结果表明具有丝素蛋白支架的新型细胞培养插入物可用作一种简便、通用且可扩展的方法来制造和运输体外 3D 共培养组织构建体，包括但不限于皮肤。由此产生的组织结构可用于治疗应用，例如作为伤口愈合中的人造皮肤替代品，以及毒理学应用，