Developmental Potential of Bovine Hand-Made Clone Embryos Reconstructed by Aggregation or Fusion with Distinct Cytoplasmic Volumes
| Autor: | Eduardo S. Ribeiro, Luciana Relly Bertolini, Maria Angélica Miglino, Fabiana Forell, Lain Uriel Ohlweiler, José Luiz Rodrigues, Joana Claudia Mezzalira, Carlos Eduardo Ambrósio, Alceu Mezzalira, Marcelo Bertolini, Ivens Ortigari, R. P. C. Gerger |
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| Rok vydání: | 2009 |
| Předmět: |
Cytoplasm
Nuclear Transfer Techniques Somatic cell Cloning Organism Parthenogenesis Embryogenesis Embryo Biology Embryo Mammalian Molecular biology Heteroplasmy In vitro medicine.anatomical_structure embryonic structures medicine Animals Cattle Blastocyst Cells Cultured Developmental Biology Biotechnology |
| Zdroj: | Cloning and Stem Cells. 11:377-386 |
| ISSN: | 1557-7457 1536-2302 |
| Popis: | Animal cloning has been associated with developmental abnormalities, with the level of heteroplasmy caused by the procedure being one of its potential limiting factors. The aim of this study was to determine the effect of the fusion of hemicytoplasts or aggregation of hemiembryos, varying the final cytoplasmic volume, on development and cell density of embryos produced by hand-made cloning (HMC), parthenogenesis or by in vitro fertilization (IVF). One or two enucleated hemicytoplasts were paired and fused with one skin somatic cell. Activated clone and zona-free parthenote embryos and hemiembryos were in vitro cultured in the well-of-the-well (WOW) system, being allocated to one of six experimental groups, on a per WOW basis: single clone or parthenote hemiembryos (1 x 50%); aggregation of two (2 x 50%), three (3 x 50%), or four (4 x 50%) clone or parthenote hemiembryos; single clone or parthenote embryos (1 x 100%); or aggregation of two clone or parthenote embryos (2 x 100%). Control zona-intact parthenote or IVF embryos were in vitro cultured in four-well dishes. Results indicated that the increase in the number of aggregated structures within each WOW was followed by a linear increase in cleavage, blastocyst rate, and cell density. The increase in cytoplasmic volume, either by fusion or by aggregation, had a positive effect on embryo development, supporting the establishment of pregnancies and the birth of a viable clone calf after transfer to recipients. However, embryo aggregation did not improve development on a hemicytoplast basis, except for the aggregation of two clone embryos. |
| Databáze: | OpenAIRE |
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