: Reproduction: oocyte enucleation

oocyte enucleation

Metaphase II oocytes are placed in HEPES-buffered HECM-2 (hamster embryo culture medium) containing 7.5 µg/ml cytochalasin B (Sigma) and incubated in this medium for 15 min prior to the start of and during enucleation. Enucleation is accomplished by using a micropipette to remove the polar body and the surrounding cytoplasm. Oocytes are enucleated while the chromosomes are being visualized to ensure successful enucleation. This involves staining the oocyte and viewing them under ultraviolet irradiation for less than 10 sec. The oocytes that are fully enucleated are returned to culture. About 38-42 h after initiation of maturation the enucleated oocytes are activated by exposure to room temperature (23-27°C) for 3 h prior to fusion. (Source : INRA)

https://opendata.inra.fr/BRMH/90

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