Animal Cap Isolation from Xenopus laevis
This protocol was adapted from “Microdissection,” Chapter 10, in Early Development of Xenopus laevis by Hazel L. Sive, Robert M. Grainger, and Richard M. Harland. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2000.INTRODUCTION
This 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.
