A modified culture method significantly improves the development of mouse somatic cell nuclear transfer embryos

Dai, Xiangpeng; Hao, Jie; Zhou, Qi

doi:10.1530/rep-09-0069

Cited by 18 publications

(12 citation statements)

References 43 publications

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“…Sorted phESCs were injected into oocytes separately to construct bimaternal embryos. The reconstructed embryos were activated by 10 mM SrCl 2 in calcium-free CZB medium at 37 C with 5% CO 2 for another 5 h. Completely activated embryos were washed and cultured in M16 medium (Sigma) at 37 C with 5% CO 2 as described (Dai et al, 2009). On the following day, bimaternal embryos at the 2-cell stage were transferred to the oviduct of psedudopregnant CD-1 mice at 0.5 dpc.…”

Section: Intracytoplasmic Injection Of Phescs and Round Spermatidmentioning

confidence: 99%

Generation of Bimaternal and Bipaternal Mice from Hypomethylated Haploid ESCs with Imprinting Region Deletions

Wang

et al. 2018

Cell Stem Cell

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Graphical Abstract Highlights d Haploid ESCs display PGC-like methylation profiles following in vitro cultivation d Parthenogenetic and androgenetic haploid ESCs show different demethylation dynamics d phESCs carrying 3 deleted imprinted regions support normal growth of bimaternal mice d ahESCs carrying 7 deleted imprinted regions produce live full-term bipaternal mice SUMMARYUnisexual reproduction is widespread among lower vertebrates, but not in mammals. Deletion of the H19 imprinted region in immature oocytes produced bimaternal mice with defective growth; however, bipaternal reproduction has not been previously achieved in mammals. We found that cultured parthenogenetic and androgenetic haploid embryonic stem cells (haESCs) display DNA hypomethylation resembling that of primordial germ cells. Through MII oocyte injection or sperm coinjection with hypomethylated haploid ESCs carrying specific imprinted region deletions, we obtained live bimaternal and bipaternal mice. Deletion of 3 imprinted regions in parthenogenetic haploid ESCs restored normal growth of fertile bimaternal mice, whereas deletion of 7 imprinted regions in androgenetic haploid ESCs enabled production of live bipaternal mice that died shortly after birth. Phenotypic analyses of organ and body size of these mice support the genetic conflict theory of genomic imprinting. Taken together, our results highlight the factors necessary for crossing same-sex reproduction barriers in mammals.

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Section: Intracytoplasmic Injection Of Phescs and Round Spermatidmentioning

confidence: 99%

Generation of Bimaternal and Bipaternal Mice from Hypomethylated Haploid ESCs with Imprinting Region Deletions

Wang

et al. 2018

Cell Stem Cell

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“…As a histone deacetylase inhibitor, CBHA could adjust acetylation sites and DNA methylation in the process of SCNT embryos' development, which was similar to that happened in zygotes. CBHA could not only increase fetus rate of cloned embryos, but also improve the derivation efficiency of NT-ES cell lines [16,32]. In this study, we found that CBHA could improve pluripotency of androgenetic embryo-derived aES cells.…”

Section: Discussionmentioning

confidence: 51%

Derivation of androgenetic embryonic stem cells from m-carboxycinnamic acid bishydroxamide (CBHA) treated androgenetic embryos

et al. 2013

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The androgenetic embyronic stem (aES) cells are useful models in studying the effects of imprinted genes on pluripotency maintaining and embryo development. The expression patterns of imprinted genes are significantly different between uniparental derived aES cells and zygote-derived embryonic stem (ES) cells, therefore, the imprinting related cell pluripotency needs further exploitation. Several approaches have been applied in generation of androgenetic embryos and derivation of aES cell lines. Here, we describe a method to generate androgenetic embryos by injecting two mature sperms into one enucleated oocyte. Then these androgenetic embryos were treated with a histone deacetylase inhibitor: m-carboxycinnamic acid bishydroxamide (CBHA). Further, aES cell lines were successfully derived from these treated androgenetic embryos at blastocyst stage. The CBHA could improve not only the quality of androgenetic embryos, but also the efficiencies of aES (CaES) cells derivation and chimeric mice generation. The imprinted gene expression pattern in the CBHA treated embryo-derived aES (CaES) cells was also highly similar to that of zygote-derived ES cells. CBHA, androgenetic, aES, pluripotency Citation:Shuai L, Feng C J, Zhang H J, et al. Derivation of androgenetic embryonic stem cells from m-carboxycinnamic acid bishydroxamide (CBHA) treated androgenetic embryos.

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“…Nuclear transfer was carried out as described . Briefly, recipient oocytes were collected from superovulated BDF1 female mice and enucleated in M2 medium (Sigma–Aldrich) containing 7.5 μg/ml cytochalasin B (Sigma–Aldrich).…”

Section: Methodsmentioning

confidence: 99%

Xist Intron 1 Repression by Transcriptional-Activator-Like Effectors Designer Transcriptional Factor Improves Somatic Cell Reprogramming in Mice

et al. 2019

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Xist is the master regulator of X chromosome inactivation. In order to further understand the Xist locus in the reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) and in somatic cell nuclear transfer (SCNT), we tested transcription-activator-like effectors-based designer transcriptional factors (dTFs), which were specific to numerous regions at the Xist locus. We report that the selected dTF repressor 6 (R6) binding the intron 1 of Xist, which caused higher H3K9me3 followed by X chromosome opening and repression of X-linked genes in mouse embryonic fibroblasts, rather than affecting Xist expression, substantially improved the iPSC generation and the SCNT preimplantation embryo development. Conversely, the dTF activator targeting the same genomic region of R6 decreased iPSC formation and blocked SCNT-embryo development. These results thus uncover the critical requirement for the Xist locus in epigenetic resetting, which is not directly related to Xist transcription. This may provide a unique route to improving the reprogramming. STEM CELLS 2019;37:599-608 SIGNIFICANCE STATEMENTUsing transcription-factor-like effectors-based designer transcriptional factors for the intron 1 of Xist, which has no effects on Xist expression, this study showed that epigenetic perturbation at the Xist locus can profoundly affect reprogramming of somatic cells, including both induced pluripotent stem cell production and somatic cell nuclear transfer-derived embryo development in vitro, which may provide a unique route to improving the reprogramming.

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A modified culture method significantly improves the development of mouse somatic cell nuclear transfer embryos

Cited by 18 publications

References 43 publications

Generation of Bimaternal and Bipaternal Mice from Hypomethylated Haploid ESCs with Imprinting Region Deletions

Generation of Bimaternal and Bipaternal Mice from Hypomethylated Haploid ESCs with Imprinting Region Deletions

Derivation of androgenetic embryonic stem cells from m-carboxycinnamic acid bishydroxamide (CBHA) treated androgenetic embryos

Xist Intron 1 Repression by Transcriptional-Activator-Like Effectors Designer Transcriptional Factor Improves Somatic Cell Reprogramming in Mice

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