Purification of avian circulating primordial germ cells by nycodenz density gradient centrifugation.
1. Primordial germ cells (PGCs) are the progenitor cells for gametes. In aves, PGCs show a unique migration pathway, that is, they circulate temporarily through the bloodstream during early development. 2. In this study we developed a method to purify circulating primordial germ cells (cPGCs) in quail and chicks by Nycodenz density gradient centrifugation. 3. The process consisted of primary and secondary purification. In primary purification, cPGCs were enriched at the interface of 8 and 12% Nycodenz fractions. In secondary purification, cPGCs were harvested from 8% Nycodenz fraction at a purity of 90% and from 10% Nycodenz fraction at a purity of 70%. The recovery rate of cPGCs was over 70%. 4. This method would facilitate research on cPGCs' culture and the production of transgenic birds using cPGCs.
Zhao
D F
DF
Department of Materials and Life Science, Graduate School of Science and Technology, Kumamoto University, Kumamoto, Japan.
Kuwana
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D000818:Animals; D002469:Cell Separation / Q000379:methods; D002499:Centrifugation, Density Gradient; D002642:Chick Embryo; D002645:Chickens; D003370:Coturnix; D005854:Germ Cells / Q000166:cytology*; D007472:Iohexol; D009024:Morphogenesis
D007472:Iohexol