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Artigo em Inglês | IMSEAR | ID: sea-38301

RESUMO

The objective of the study was to develop the somatic nuclear transfer technique by using rabbits as the model. The oocyte recipients aged 16 h post coitus were collected surgically from 20 superovulated rabbit doe with 28 and 40 mg Follicle Stimulating Hormone (FSH) after mating with a vasectomized male. The metaphase II plate and 1st polar body of oocyte was later aspirated by enucleated micropipette under an inverted microscope. A single donor cell; cumulus cell or cultured or frozen fibroblast cell from passage 1 to 9 were transferred to enucleated oocyte and fused with triple DC pulses, 3.2 kv, 20 micros. The fused embryos were cultivated in TCM 199 NaHCO3 + 10 per cent fetal calf serum (FCS) for 4 days. The cleavage rate (2-cell stage) was 37.2 per cent (32/86) from eight experiments, and 18.8 per cent (6/32) developed to the early morula stage. This study also indicated that the enucleation pipette and the somatic cell type influenced the success.


Assuntos
Transporte Ativo do Núcleo Celular , Animais , Clonagem de Organismos/métodos , Técnicas de Cocultura , Desenvolvimento Embrionário e Fetal/fisiologia , Feminino , Fibroblastos/citologia , Masculino , Modelos Animais , Oócitos/crescimento & desenvolvimento , Coelhos , Sensibilidade e Especificidade
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