Use of retroviral vectors to introduce and express the beta-galactosidase marker gene in cultured chicken primordial germ cells.
Three methods of isolating primordial germ cells (PGCs) from gonads of 5-day-old chick embryos were compared. PGCs were then cultured in vitro in DMEM/F12 medium containing 10% fetal calf serum. BrdU incorporation showed that at least 10% of the PGC population were dividing, under our culture conditions, during the 2nd day of in vitro culture. During this culture period, PGCs were exposed to avian leukosis sarcoma virus-based retroviral vector pseudotyped with subgroup A envelope, carrying the LacZ reporter gene. X-Gal staining showed that PGCs were permissive to infection, with more than 50% of PGCs expressing the beta-Gal protein. These data represent the first demonstration that PGCs, isolated from gonads of 5-day-old chick embryos, are able to divide in vitro and that it is possible to introduce and express exogenous DNA in chick PGCs maintained in vitro.
Allioli
N
N
Centre de Génétique Moléculaire et Cellulaire, CNRS UMR106, INRA LA810, Université Claude Bernard Lyon-I, Villeurbanne, France.
Thomas
J L
JL
Chebloune
Y
Y
Nigon
V M
VM
Verdier
G
G
Legras
C
C
D000818:Animals; D001354:Avian Leukosis Virus / Q000235:genetics*; D002455:Cell Division / Q000235:genetics; D002478:Cells, Cultured; D002642:Chick Embryo; D005819:Genetic Markers; D005822:Genetic Vectors; D005854:Germ Cells / Q000378:metabolism*; D006066:Gonads / Q000196:embryology; D001616:beta-Galactosidase / Q000235:genetics*
D005819:Genetic Markers; D001616:beta-Galactosidase