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Purification of primordial germ cells from TNAPbeta-geo mouse embryos using FACS-gal
K Abe1, M Hashiyama, G Macgregor
1Institute of Molecular Embryology and Genetics, Kumamoto University School of Medicine, 4-24-1 Kuhonji, Kumamoto, 862, Japan. kabe@gpo.kumamoto-u.ac.jp
Abstract:
Tissue nonspecific alkaline phosphatase (TNAP), the product of the Akp2 locus, is expressed in mouse primordial germ cells (PGC) for an extensive period during embryogenesis. Mice with the Akp2tm1Sor mutant allele of TNAP express lacZ (beta-galactosidase; beta-gal) under control of the Akp2 locus. PGCs were purified from Akp2tm1Sor embryos using fluorescence activated cell sorting of beta-gal expressing cells (FACS-gal). Analysis of the purified cells by alkaline phosphatase staining and immunocytochemistry with anti-c-kit antibody demonstrated that highly (98%) purified PGCs can be isolated using this method. This technique will facilitate experiments that require highly purified preparations of PGCs including cell culture and gene expression analyses.