Production of bovine hand-made cloned embryos by zygote-oocyte cytoplasmic hemi-complementation
| Autor: | Carlos Eduardo Ambrósio, Marcelo Bertolini, Lain Uriel Ohlweiler, R. P. C. Gerger, Maria Angélica Miglino, R. Casali, Joana Claudia Mezzalira, Alceu Mezzalira, José Luiz Rodrigues, F. K. Vieira |
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| Rok vydání: | 2011 |
| Předmět: |
endocrine system
Cytoplasm Nuclear Transfer Techniques Zygote Cloning Organism Parthenogenesis Fertilization in Vitro Biology Cytoplast Membrane Fusion EMBRIÃO DE ANIMAL Andrology medicine Animals Blastocyst reproductive and urinary physiology Cloning Genetics urogenital system Embryo Cell Biology Oocyte female genital diseases and pregnancy complications Complementation medicine.anatomical_structure embryonic structures Oocytes Cattle Female Developmental Biology Biotechnology |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| ISSN: | 2152-4998 |
| Popis: | The aim of this study was to evaluate the effect of the cytoplast type and activation process on development of cloned embryos. Bovine oocytes (MII) or zygotes at the one-cell stage (IVF) were manually bisected and segregated in MII or IVF hemi-cytoplasts or hemi-karyoplasts. Adult skin cells from a bovine female were used as nucleus donors (SC). Experimental groups were composed of IVF embryos; parthenogenetic embryos; hand-made cloned (HMC) embryos; and reconstructed HMC embryos using IVF hemi-cytoplast + MII hemi-cytoplast + SC (G-I); IVF hemi-cytoplast + IVF hemi-cytoplast + SC (G-II); MII hemi-cytoplast + IVF hemi-karyoplast (G-III); and IVF hemi-cytoplast + IVF hemi-karyoplast (G-IV). Embryos from G-I to G-IV were allocated to subgroups as sperm-activated (SA) or were further chemically activated (SA + CA). Embryos from all groups and subgroups were in vitro cultured in the WOW system. Blastocyst development in subgroup G-I SA (28.2%) was similar to IVF (27.0%) and HMC (31.4%) controls, perhaps due to a to a more suitable activation process and/or better complementation of cytoplasmic reprogramming factors, with the other groups and subgroups having lower levels of development. No blastocyst development was observed when using IVF hemi-karyoplasts (G-III and G-IV), possibly due to the manipulation process during a sensitive biological period. In summary, the presence of cytoplasmic factors from MII hemi-oocytes and the sperm activation process from hemi-zygotes appear to be necessary for adequate in vitro development, as only the zygote-oocyte hemi-complementation was as efficient as controls for the generation of bovine cloned blastocysts. |
| Databáze: | OpenAIRE |
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