Immunomagnetic purification of murine primordial germ cells.
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.
Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL, USA.
Emily Y
EY
Smith
James L
JL
Resnick
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D047468:Immunoprecipitation / Q000379:methods*; D060526:Magnetic Fields; D051379:Mice; D008863:Microspheres; D010063:Ovum / Q000166:cytology*
D000469:Alkaline Phosphatase