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 vit ro in DMEM/F12 medium containing 10% fetal calf serum. BrdU incorporat ion showed that at least 10% of the PGC population were dividing, unde r our culture conditions, during the 2nd day of in vitro culture. Duri ng this culture period, PGCs were exposed to avian leukosis sarcoma vi rus-based retroviral vector pseudotyped with subgroup A envelope, carr ying the LacZ reporter gene. X-Gal staining showed that PGCs were perm issive to infection, with more than 50% of PGCs expressing the beta-Ga l protein. These data represent the first demonstration that PGCs, iso lated from gonads of 5-day-old chick embryos, are able to divide in vi tro and that it is possible to introduce and express exogenous DNA in chick PGCs maintained in vitro. (C) 1994 Academic Press, Inc.