Ishida Kentaro, Mitsui Toshiyuki
Department of Physics and Mathematics, College of Science and Engineering, Aoyama Gakuin University, Kanagawa, 252-5258, Japan.
Dev Growth Differ. 2016 Apr;58(3):303-14. doi: 10.1111/dgd.12275. Epub 2016 Mar 28.
Various kinds of in vitro culture systems of tissues and organs have been developed, and applied to understand multicellular systems during embryonic organogenesis. In the research field of feather bud development, tissue recombination assays using an intact epithelial tissue and mesenchymal tissue/cells have contributed to our understanding the mechanisms of feather bud formation and development. However, there are few methods to generate a skin and its appendages from single cells of both epithelium and mesenchyme. In this study, we have developed a bioengineering method to reconstruct an embryonic dorsal skin after completely dissociating single epithelial and mesenchymal cells from chick skin. Multiple feather buds can form on the reconstructed skin in a single row in vitro. The bioengineered feather buds develop into long feather buds by transplantation onto a chorioallantoic membrane. The bioengineered bud sizes were similar to those of native embryo. The number of bioengineered buds was increased linearly with the initial contact length of epithelial and mesenchymal cell layers where the epithelial-mesenchymal interactions occur. In addition, the bioengineered bud formation was also disturbed by the inhibition of major signaling pathways including FGF (fibroblast growth factor), Wnt/β-catenin, Notch and BMP (bone morphogenetic protein). We expect that our bioengineering technique will motivate further extensive research on multicellular developmental systems, such as the formation and sizing of cutaneous appendages, and their regulatory mechanisms.
已经开发出各种组织和器官的体外培养系统，并将其应用于了解胚胎器官发生过程中的多细胞系统。在羽毛芽发育的研究领域中，使用完整上皮组织和间充质组织/细胞的组织重组试验有助于我们理解羽毛芽形成和发育的机制。然而，从上皮和间充质的单细胞生成皮肤及其附属器的方法很少。在本研究中，我们开发了一种生物工程方法，在将鸡皮肤的单个上皮细胞和间充质细胞完全解离后重建胚胎背部皮肤。多个羽毛芽可以在体外重建的皮肤上单行形成。通过移植到尿囊膜上，生物工程化的羽毛芽发育成长羽毛芽。生物工程化芽的大小与天然胚胎的相似。生物工程化芽的数量与发生上皮-间充质相互作用的上皮和间充质细胞层的初始接触长度呈线性增加。此外，生物工程化芽的形成也受到包括FGF（成纤维细胞生长因子）、Wnt/β-连环蛋白、Notch和BMP（骨形态发生蛋白）在内的主要信号通路抑制的干扰。我们期望我们的生物工程技术将推动对多细胞发育系统的进一步广泛研究，例如皮肤附属器的形成和大小以及它们的调节机制。
