PURIFICATION OF PRIMORDIAL GERM-CELLS FROM TNAP(BETA-GEO) MOUSE EMBRYOS USING FACS-GAL

Tissue nonspecific alkaline phosphatase (TNAP), the product of the Akp 2 locus, is expressed in mouse primordial germ cells (PGC) for an exte nsive period during embryogenesis. Mice with the Akp2(tm/Sor) mutant a llele of TNAP express lacZ (beta-galactosidase; beta-gal) under contro l of the Akp2 locus. PGCs were purified from Akp2(tm/Sor) embryos usin g fluorescence activated cell sorting of beta-gal expressing cells (FA CS-gal), Analysis of the purified cells by alkaline phosphatase staini ng and immunocytochemistry with anti-c-kit antibody demonstrated that highly (98%) purified PGCs can be isolated using this method. This tec hnique will facilitate experiments that require highly purified prepar ations of PGCs including cell culture and gene expression analyses. (C ) 1996 Academic Press, Inc.