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• W2007356572 abstract "Reprogramming after somatic cell nuclear transfer had been thought to be dependent on the recipient cytoplasm being arrested at the metaphase stage, but here interphase two-cell mouse embryos are shown to support successful reprogramming and generation of embryonic stem cells or cloned mice. The reprogramming that occurs following somatic cell nuclear transfer (SCNT) during infertility treatment and other procedures is thought to be dependent on the recipient cytoplasm being strictly arrested at metaphase. Shoukhrat Mitalipov and colleagues now show that interphase two-cell mouse embryos support the reprogramming of transplanted somatic nuclei and generation of embryonic stem cells or cloned mice. This suggests that factors capable of inducing pluripotency are present in the cytoplasm of interphase cells, and that removal of the nuclei of such cells does not deplete a recipient egg of necessary reprogramming factors as was thought. If these findings extend to humans, they could boost efforts to generate human embryonic stem cells for regenerative applications, as donated cells in the interphase stage (fertilized embryos) are more accessible than those in the metaphase stage (immature egg cells). Successful mammalian cloning using somatic cell nuclear transfer (SCNT) into unfertilized, metaphase II (MII)-arrested oocytes attests to the cytoplasmic presence of reprogramming factors capable of inducing totipotency in somatic cell nuclei1,2,3. However, these poorly defined maternal factors presumably decline sharply after fertilization, as the cytoplasm of pronuclear-stage zygotes is reportedly inactive4,5. Recent evidence suggests that zygotic cytoplasm, if maintained at metaphase, can also support derivation of embryonic stem (ES) cells after SCNT6,7,8, albeit at low efficiency. This led to the conclusion that critical oocyte reprogramming factors present in the metaphase but not in the interphase cytoplasm are ‘trapped’ inside the nucleus during interphase and effectively removed during enucleation9. Here we investigated the presence of reprogramming activity in the cytoplasm of interphase two-cell mouse embryos (I2C). First, the presence of candidate reprogramming factors was documented in both intact and enucleated metaphase and interphase zygotes and two-cell embryos. Consequently, enucleation did not provide a likely explanation for the inability of interphase cytoplasm to induce reprogramming. Second, when we carefully synchronized the cell cycle stage between the transplanted nucleus (ES cell, fetal fibroblast or terminally differentiated cumulus cell) and the recipient I2C cytoplasm, the reconstructed SCNT embryos developed into blastocysts and ES cells capable of contributing to traditional germline and tetraploid chimaeras. Last, direct transfer of cloned embryos, reconstructed with ES cell nuclei, into recipients resulted in live offspring. Thus, the cytoplasm of I2C supports efficient reprogramming, with cell cycle synchronization between the donor nucleus and recipient cytoplasm as the most critical parameter determining success. The ability to use interphase cytoplasm in SCNT could aid efforts to generate autologous human ES cells for regenerative applications, as donated or discarded embryos are more accessible than unfertilized MII oocytes." @default.
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• W2007356572 date "2014-03-26" @default.
• W2007356572 modified "2023-09-27" @default.
• W2007356572 title "Nuclear reprogramming by interphase cytoplasm of two-cell mouse embryos" @default.
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• W2007356572 doi "https://doi.org/10.1038/nature13134" @default.
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