Nuclear reprogramming by interphase cytoplasm of two-cell mouse embryos.
Nuclear reprogramming by interphase cytoplasm of two-cell mouse embryos.
复制标题
DOI：
10.1038/nature13134
复制
发表时间：
2014-05-01
期刊：
影响因子：
64.8
通讯作者：
Mitalipov, Shoukhrat
中科院分区：
文献类型：
--
作者：
Kang, Eunju;Wu, Guangming;Ma, Hong;Li, Ying;Tippner-Hedges, Rebecca;Tachibana, Masahito;Sparman, Michelle;Wolf, Don P.;Schoeler, Hans R.;Mitalipov, Shoukhrat
利用体细胞核移植(SCNT)成功地将哺乳动物克隆到未受精的中期II停滞(MII)卵母细胞中，证明了胞质中存在能够诱导体细胞核多能性的重编程因子。然而，这些定义不明确的母体因子可能在受精后急剧下降，因为据报道，原核期受精卵的细胞质不活跃。最近的证据表明，如果合子细胞质保持在中期(M期)，也可以支持SCNT后胚胎干细胞(ESCs)的起源，尽管效率较低。这导致的结论是，关键的卵母细胞重编程因子存在于M期，但不存在于间期细胞质中，在间期被困在核内，并在去核过程中有效地被移除。在这里，我们研究了2-细胞小鼠胚胎(I2C)间期细胞质中重编程活性的存在。首先，在完整的和去核的M期和间期受精卵和2-细胞胚胎中都存在候选重编程因子。因此，去核不能提供间期细胞质不能诱导重编程的可能解释。然后，当我们仔细地将移植核(ESC、胎儿成纤维细胞或终末分化的卵丘细胞)和受体I2C细胞质之间的细胞周期阶段同步时，重构的SCNT胚胎发育成囊胚和ESCs，能够促进传统的种系和四倍体嵌合体的形成。此外，将用胚胎干细胞核重建的克隆胚胎直接移植到受体体内，会产生活的后代。因此，I2C的细胞质支持有效的重编程，供体核和受体细胞质之间的细胞周期同步是决定成功的最关键参数。在SCNT中利用间期细胞质的能力可能会影响产生用于再生应用的自体人类ESCs的努力，因为捐赠或丢弃的胚胎比未受精的MII卵母细胞更容易获得。
Successful mammalian cloning employing somatic cell nuclear transfer (SCNT) into unfertilized, metaphase II-arrested (MII) oocytes attests to the cytoplasmic presence of reprogramming factors capable of inducing pluripotency in somatic cell nuclei. However, these poorly defined maternal factors presumably decline sharply after fertilization since cytoplasm of pronuclear stage zygotes is reportedly inactive. Recent evidence suggests that zygotic cytoplasm, if maintained at metaphase (M-phase) can also support derivation of embryonic stem cells (ESCs) following SCNT, albeit at low efficiency. This led to the conclusion that critical oocyte reprogramming factors present in M-phase but not in interphase cytoplasm are “trapped” inside the nucleus during interphase and effectively removed during enucleation. Here, we investigated the presence of reprogramming activity in the interphase cytoplasm of 2-cell mouse embryos (I2C). First, the presence of candidate reprogramming factors was documented in both intact and enucleated M-phase and interphase zygotes and 2-cell embryos. Consequently, enucleation did not provide a likely explanation for the inability of interphase cytoplasm to induce reprogramming. Then, when we carefully synchronized the cell cycle stage between the transplanted nucleus (ESC, fetal fibroblast or terminally differentiated cumulus cell) and the recipient I2C cytoplasm, the reconstructed SCNT embryos developed into blastocysts and ESCs capable of contributing to traditional germline and tetraploid chimeras. In addition, direct transfer of cloned embryos, reconstructed with ESC 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 utilize interphase cytoplasm in SCNT could impact efforts to generate autologous human ESCs for regenerative applications since donated or discarded embryos are more accessible than unfertilized, MII oocytes.
登录
查看更多内容
影响因子：
8.8
作者：
Lee HS;Ma H;Juanes RC;Tachibana M;Sparman M;Woodward J;Ramsey C;Xu J;Kang EJ;Amato P;Mair G;Steinborn R;Mitalipov S
通讯作者：
Mitalipov S
影响因子：
--
作者：
TSUNODA, Y;YASUI, T;SUGIE, T
通讯作者：
SUGIE, T
DOI：
10.1073/pnas.96.26.14984
发表时间：
1999-12-21
影响因子：
11.1
作者：
Wakayama, T;Rodriguez, I;Mombaerts, P
通讯作者：
Mombaerts, P
影响因子：
6.1
作者：
Mitalipov, S. M.;Zhou, Q.;Wolf, D. P.
通讯作者：
Wolf, D. P.
影响因子：
9.8
作者：
Wu G;Liu N;Rittelmeyer I;Sharma AD;Sgodda M;Zaehres H;Bleidissel M;Greber B;Gentile L;Han DW;Rudolph C;Steinemann D;Schambach A;Ott M;Schöler HR;Cantz T
通讯作者：
Cantz T