Simple separation of chicken gonadal primordial germ cells with and without foreign genes

Abstract

Simplicity is the key element of an inexpensive technique described that is superior in performance to previous methods. It can make it the rapid method of choice to obtain reasonable yields of purified primordial germ cells (PGCs) for immediate production of germline chimeric chickens with integrated foreign genes. After Ficoll centrifugation, the purity of PGCs from gonads was 80.9 0.08% (mechanical) compared with 86.1 0.19% (enzymatic). GFP gene and lacZtransduced chicken gonadal primordial germ cells (gPGCs) examined 72 h after transduction had a transfection efficiency of 61% and 64%, respectively. After 10 days of G418 selection, approximately 90 and 92% of pure gPGCs did not contain other cells following this Ficoll gradient centrifugation method of preparation.