Culturing in vitro produced blastocysts in sequential media promotes ES cell derivation.
Mol Reprod Dev
; 73(8): 1017-21, 2006 Aug.
Article
en En
| MEDLINE
| ID: mdl-16700070
Embryonic stem (ES) cell lines are routinely derived from in vivo produced blastocysts. We investigated the efficiency of ES cells derivation from in vitro produced blastocysts either in monoculture or sequential culture. Zygotes from hybrid F1 B6D2 mice were cultured in vitro to the blastocyst stage in Potassium (K(+)) simplex optimised medium (KSOM) throughout or in KSOM and switched to COOK blastocyst medium on day 3 (KSOM-CBM). Blastocysts were explanted on a feeder layer of mitomycin C-inactivated murine embryonic fibroblasts (MEF) in TX-WES medium for ES cell derivation. Sequential KSOM-CBM resulted in improved blastocyst formation compared to KSOM monoculture. ES cells were obtained from 32.1% of explanted blastocsyts cultured in KSOM-CBM versus 18.4% in KSOM alone. ES cell lines were characterized by morphology, expression of SSEA-1, Oct-4 and alkaline phosphatase activity, and normal karyotype. These results indicate that in vitro culture systems to produce blastocysts can influence the efficiency of ES cell line derivation.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Blastocisto
/
Técnicas de Cultivo de Célula
/
Células Madre Embrionarias
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Mol Reprod Dev
Asunto de la revista:
BIOLOGIA MOLECULAR
/
MEDICINA REPRODUTIVA
Año:
2006
Tipo del documento:
Article
País de afiliación:
Bélgica
Pais de publicación:
Estados Unidos