Ariizumi Takashi, Michiue Tatsuo, Asashima Makoto
Department of Agri-Production Sciences, Tamagawa University, Machida, Tokyo 194-8610, Japan.
Department of Life Sciences (Biology), Graduate School of Arts and Sciences, The University of Tokyo, Meguro, Tokyo 153-8902, Japan.
Cold Spring Harb Protoc. 2017 Dec 1;2017(12):pdb.prot097410. doi: 10.1101/pdb.prot097410.
The animal cap-the presumptive ectoderm of the blastula embryo-can differentiate into a variety of tissues belonging to the three germ layers following exposure to specific inducers. The "animal cap assay" was devised based on the pluripotency of presumptive ectodermal cells and enabled many important discoveries in the field of embryonic induction and cell differentiation. Using this system, investigators can test multiple factors in solution simultaneously to determine their inducing activities qualitatively, quantitatively, and synergistically. Furthermore, after dissociation and induction, reaggregated animal cap cells can be induced to form higher-order organs. This protocol details preoperative preparations, followed by the basic animal cap assay. Advanced protocols for the induction of kidney, pancreas, and heart are also described.
动物帽——囊胚胚胎的预定外胚层——在暴露于特定诱导剂后可分化为属于三个胚层的多种组织。“动物帽实验”是基于预定外胚层细胞的多能性设计的，它在胚胎诱导和细胞分化领域促成了许多重要发现。利用这个系统，研究人员可以同时测试溶液中的多种因子，以定性、定量和协同地确定它们的诱导活性。此外，在解离和诱导后，重新聚集的动物帽细胞可以被诱导形成高阶器官。本方案详细介绍了术前准备，然后是基本的动物帽实验。还描述了诱导肾脏、胰腺和心脏的进阶方案。
