Mature MII oocytes were recovered from females undergoing controlled ovarian stimulations. Oocytes were transferred to a manipulation chamber containing medium with 5 μg ml cytochalasin B and MII spindle-chromosomal complexes were aspirated and isolated using Oosight imaging system (www.cri-inc.com ) as described before16 (link). Isolated karyoplasts were briefly exposed to SeV extract (www.cosmobio.co.jp ) and placed into perivitelline space on the side opposite to the 1st polar body (Suppl. video). During manipulations, oocyte's zona pellucida was penetrated by laser assisted zona drilling (www.hamiltonthorne.com ). After fusion, reconstructed oocytes were fertilized by ISCI and cultured as described before16 (link). Detailed methods are described in Supplementary information at www.nature.com/nature .
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Nuclear Transfer in Mature Oocytes
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Other organizations : Oregon National Primate Research Center
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Variable analysis
independent variables
- Removal of MII spindle-chromosomal complexes using Oosight imaging system
- Exposure of isolated karyoplasts to SeV extract
- Penetration of oocyte's zona pellucida by laser assisted zona drilling
- Fusion of reconstructed oocytes
- Fertilization of reconstructed oocytes by ICSI
- Subsequent culture of reconstructed oocytes
- Mature MII oocytes recovered from females undergoing controlled ovarian stimulation
- Oocytes transferred to a manipulation chamber containing medium with 5 μg ml cytochalasin B
- Placement of isolated karyoplasts into perivitelline space on the side opposite to the 1st polar body
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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