Production of transgenic recloned piglets harboring the human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene from porcine fetal fibroblasts by nuclear transfer.
Theriogenology
; 70(9): 1431-8, 2008 Dec.
Article
em En
| MEDLINE
| ID: mdl-18804273
ABSTRACT
We used nuclear transfer (NT) to develop transgenic female pigs harboring goat beta-casein promoter/human granulocyte-macrophage colony stimulating factor (hGM-CSF). The expression of hGM-CSF was specific to the mammary gland, and the glycosylation-derived size heterogeneity corresponded to that of the native human protein. Although various cell types have been used to generate cloned animals, little is currently known about the potential use of fibroblasts derived from a cloned fetus as donor cells for nuclear transfer. The developmental potential of porcine cloned fetal fibroblasts transfected with hGM-CSF was evaluated in the present study. Cloned fetal fibroblasts were isolated from a recipient following the transplantation of NT embryos. The cells were transfected with both hGM-CSF and the neomycin resistance gene in order to be used as donor cells for NT. Reconstructed embryos were implanted into six sows during estrus; two of the recipient sows delivered seven healthy female piglets with the hGM-CSF gene (confirmed with PCR and fluorescent in situ hybridization) and microsatellite analysis confirmed that the clones were genetically identical to the donor cells. The expression of hGM-CSF was strong in the mammary glands of a transgenic pig that died a few days prior to parturition (110 d after AI). These results demonstrated that somatic cells derived from a cloned fetus can be used to produce recloned and transgenic pigs.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Suínos
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Animais Geneticamente Modificados
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Fator Estimulador de Colônias de Granulócitos e Macrófagos
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Técnicas de Transferência Nuclear
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Fibroblastos
Limite:
Animals
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Female
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Humans
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Pregnancy
Idioma:
En
Revista:
Theriogenology
Ano de publicação:
2008
Tipo de documento:
Article