Use of retroviral vectors to introduce and express the beta-galactosidase marker gene in cultured chicken primordial germ cells.
Dev Biol
; 165(1): 30-7, 1994 Sep.
Article
en En
| MEDLINE
| ID: mdl-8088448
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.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Marcadores Genéticos
/
Beta-Galactosidasa
/
Virus de la Leucosis Aviar
/
Vectores Genéticos
/
Células Germinativas
Límite:
Animals
Idioma:
En
Revista:
Dev Biol
Año:
1994
Tipo del documento:
Article
País de afiliación:
Francia
Pais de publicación:
Estados Unidos