Sive Hazel L, Grainger Robert M, Harland Richard M
CSH Protoc. 2007 Jun 1;2007:pdb.prot4744. doi: 10.1101/pdb.prot4744.
INTRODUCTIONThis protocol presents a method for isolating Xenopus laevis animal cap cells, which are cells situated around the animal (pigmented) pole of a blastula or very early gastrula-stage embryo. This tissue is fated to become cement gland/neurectoderm on the dorsal side of the embryo and epidermis on the ventral side. Animal caps are composed of pluripotent cells that can be induced to form endodermal, mesodermal, or ectodermal cell types, and can therefore serve as a useful substrate to assess the activity of various inducing factors. Caps from embryos that have been injected with expression constructs can also be removed and analyzed to assess the activity of various genes. In addition, caps can be used in conjugation (induction) assays.
引言
本方案介绍了一种分离非洲爪蟾动物帽细胞的方法，这些细胞位于囊胚或极早期原肠胚阶段胚胎的动物（色素沉着）极周围。该组织注定会在胚胎的背侧形成黏着腺/神经外胚层，在腹侧形成表皮。动物帽由多能细胞组成，这些细胞可被诱导形成内胚层、中胚层或外胚层细胞类型，因此可作为评估各种诱导因子活性的有用底物。已注射表达构建体的胚胎的帽也可被移除并进行分析，以评估各种基因的活性。此外，动物帽可用于结合（诱导）试验。
