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- W2360670383 abstract "Induced enucleation is carried out with agents disturbing the meiosis or inhibiting protein synthesis of oocyte during polar body emites.Consequently all chromatin are extruded as the form of polar body.The mice have been cloned from embryonic stem cell karyoplasts and activated cytoplasts prepared by induced enucleation of MⅡ oocytes.However the nuclear transfer of mice with the induced-enucleation MⅡ oocytes as the recipient has not been reported.Compared to the traditionally micromanipulation,the induced enucleation with chemical does not damage the cytoplast cytoskeleton,reduce the volume and cause the lost of some important factors involved in reprogramming of donor karyoplast.Moreover its high efficiency,time-saving and simple procedures maybe make the cytoplast derived from the induced enucleation more suitable to the reprogramming of donor nuclear.Ovaries were removed from animals and immersed in M2.The GV oocytes were collected by puncturing the large antral follicies using 25-gauge needles and matured in vitro.After 5 h IVM,oocytes that had undergone GVBD were cultured in M16 containing DC for 2 h,then in M16 with DC and CHX sequently until the first polar body was extruded.Completely enucleated oocytes were transferred into the M2 medium with 0.5% protease to remove the zona.Then the mouse fetal fibroblast cells were glued to the oocytes membrane with PHA.Consequently the oocyte-cell couplets were fused by electronic pulse and the fused couplets were transfer into medium with SrCl_2 for 6 h.Finally the reconstructed embryos were cultured in well of well with M16.In total,698 oocytes were treated in our experiment with three repeats.The rates of fused and activated reconstruction embryos were 84.8% and 93.6% respetively. Embryos in 4-cell stage were derived by this method.No pregnancy was observed after the 2-cell embryos were transferred to the surrogate mice.Additionally,the effects of 3 protocols for donor preparation(trypsinizing the cultured cells with or without serum starvation,freeze preservation of cultured cells)on the development of reconstructed embryos were studied.The results showed there were no significant differences (P0.05) among groups except fusion rate(P0.05).And thus the freeze preservation is a simple but efficacious protocol for preparing donor cells in nuclear transfer.In generally,we combined the induced enucleation and the zona-free technology in mouse somatic cell nuclear transfer.Surely further research is required to improve the developmental potential of reconstructed embryos produced by this new method and its availability will greatly facilitate the procedure and improve the total efficiency of nuclear transfer in mammal." @default.
- W2360670383 created "2016-06-24" @default.
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- W2360670383 date "2006-01-01" @default.
- W2360670383 modified "2023-09-25" @default.
- W2360670383 title "Reconstruction of mouse embryos with chemically enucleated oocytes" @default.
- W2360670383 hasPublicationYear "2006" @default.
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