Smith Emily Y, Resnick James L
Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL, USA.
Methods Mol Biol. 2012;925:61-6. doi: 10.1007/978-1-62703-011-3_4.
Primordial germ cells (PGCs) play essential roles in both reproduction and development. In this chapter, we describe a method used in our laboratory for the immunopurification of PGCs from the mouse embryo. After dissection and disruption of the fetal gonad, PGCs are identified by a monoclonal antibody recognizing an epitope characteristic of pluripotent stem cells. After reaction with a paramagnetic bead-linked secondary antibody, the cell mixture is applied to a strong magnetic field. PGCs are recovered by release from the magnetic field. Purity is assessed by the alkaline phosphatase activity inherent to PGCs.
