Simple separation of chicken gonadal primordial germ cells with and without foreign genes.
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 lacZ-transduced chicken gonadal primordial germ cells (gPGCs) examined 72h after transduction had a transfection efficiency of approximately 61% and approximately 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.
Department of Animal Science and Technology, College of Agriculture and Life Sciences, Seoul National University, Suweon, 441-744, Korea.
Dong Kee
DK
Jeong
Yeong Ho
YH
Hong
Jae Yong
JY
Han
D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology* / Q000235:genetics; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002642:Chick Embryo / Q000166:cytology* / Q000196:embryology; D002678:Chimera / Q000235:genetics*; D005362:Ficoll; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D005854:Germ Cells / Q000166:cytology*; D006066:Gonads / Q000166:cytology* / Q000196:embryology; D019076:Transgenes / Q000235:genetics
D005362:Ficoll