Cell type-specific reporter transgenic chicken lines are invaluable tools in developmental biology, allowing the visualization of dynamics and differentiation states of target cell types in living embryos. Here, we report the establishment of a new transgenic chicken line in which limb mesenchyme and apical ectodermal ridge (AER) cells are labeled with different fluorescent proteins in the embryos. The processes for generating the reporter line involved using tissue-specific promoters, the Tol2 transposon-mediated genomic integration, and clonal culture system of primordial germ cells. Employing the transgenic chickens would facilitate the detailed characterization of limb mesenchyme and AER cells. Thus, this reporter chicken line will be a powerful tool for advancing the study of vertebrate limb development.