Mice cloned from embryonic stem cells

Wakayama, Teruhiko; Rodríguez, Iván; Perry, Anthony C.F.; Yanagimachi, Ryuzo; Mombaerts, Peter

doi:10.1073/pnas.96.26.14984

Cited by 436 publications

(285 citation statements)

References 37 publications

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“…The neonatal weight of the live pup was in the normal range (1.601 g), while the weight of the placenta was approximately three times higher (0.327 g), similar to what was observed in previous studies of cloned mice [12] ( Figure 1L). In contrast, neither pups nor degenerating embryos, but only placentas, were recovered from the tetraploid control as expected ( Figure 1K).…”

Section: Mitosis-arrested Tetraploid Embryos Support Nuclear Reprograsupporting

confidence: 75%

Mouse cloning and somatic cell reprogramming using electrofused blastomeres

et al. 2010

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Mouse cloning from fertilized eggs can assist development of approaches for the production of "genetically tailored" human embryonic stem (ES) cell lines that are not constrained by the limitations of oocyte availability. However, to date only zygotes have been successfully used as recipients of nuclei from terminally differentiated somatic cell donors leading to ES cell lines. In fertility clinics, embryos of advanced embryonic stages are usually stored for future use, but their ability to support the derivation of ES cell lines via somatic nuclear transfer has not yet been proved. Here, we report that two-cell stage electrofused mouse embryos, arrested in mitosis, can support developmental reprogramming of nuclei from donor cells ranging from blastomeres to somatic cells. Live, full-term cloned pups from embryonic donors, as well as pluripotent ES cell lines from embryonic or somatic donors, were successfully generated from these reconstructed embryos. Advanced stage pre-implantation embryos were unable to develop normally to term after electrofusion and transfer of a somatic cell nucleus, indicating that discarded pre-implantation human embryos could be an important resource for research that minimizes the ethical concerns for human therapeutic cloning. Our approach provides an attractive and practical alternative to therapeutic cloning using donated oocytes for the generation of patient-specific human ES cell lines.

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Section: Mitosis-arrested Tetraploid Embryos Support Nuclear Reprograsupporting

confidence: 75%

Mouse cloning and somatic cell reprogramming using electrofused blastomeres

et al. 2010

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“…Enucleated B6D2 F 1 oocytes were injected individually with an adult tail-tip, cumulus cell, or ntES cell nuclei (10,11,18,25), activated by using Sr 2ϩ (23), and allowed to develop to two to eight cell embryos, morulae, or blastocysts before they were transferred to pseudopregnant ICR surrogate mothers.…”

Section: Production Of Cloned Offspring By Using Adult Somatic Cells mentioning

confidence: 99%

Propagation of an infertile hermaphrodite mouse lacking germ cells by using nuclear transfer and embryonic stem cell technology

Wakayama

Kishigami

Thuan

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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Animals generated by systematic mutagenesis and routine breeding are often infertile because they lack germ cells, and maintenance of such lines of animals has been impossible. We found a hermaphrodite infertile mouse in our colony, a genetic male with an abnormal Y chromosome lacking developing germ cells. We tried to clone this mouse by conventional nuclear transfer but without success. ES cells produced from blastocysts, which had been cloned by using somatic cell nuclear transfer (ntES cells) from this mouse, were also unable to produce offspring when injected into enucleated oocytes. Although we were able to produce two chimeric offspring using these ntES cells by tetraploid complementation, they were infertile, because they also lacked developing germ cells. However, when such ntES cells were injected into normal diploid blastocysts, many chimeric offspring were produced. One such male offspring transmitted hermaphrodite mouse genes to fertile daughters via X chromosome-bearing sperm. Thus, ntES cells were used to propagate offspring from infertile mice lacking germ cells.

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“…The use of blastomeres as donor cells were successfully used to generate the first mammalian clones, [1][2][3] Embryonic stem cells have been successfully used as donor cells in the mouse. 4,5 Higher rates of development have been reported when using ES cells as the donor cells compared to somatic donor cells. 6,7 Conversely, the use of inbred 129 ES cells failed to produce any offspring that survived more than one day.…”

Section: Introductionmentioning

confidence: 99%

Correlation of Developmental Differences of Nuclear Transfer Embryos Cells to the Methylation Profiles of Nuclear Transfer Donor Cells in Swine

Bonk¹,

Cheong²,

Li³

et al. 2007

Epigenetics

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Methylation of DNA is the most commonly studied epigenetic mechanism of developmental competence and somatic cell nuclear transfer (SCNT). Previous studies of epigenetics and the SCNT procedures have examined the effects of different culture media on donor cells and reconstructed embryos, and the methylation status of specific genes in the fetus or live offspring. Here we used a microarray based approach to identify the methylation profiles of SCNT donor cells including three clonal porcine fetal fibroblast-like cell sublines and adult somatic cells selected from kidney and mammary tissues. The methylation profiles of the donor cells were then analyzed with respect to their ability to direct development to the blastocyst stage after nuclear transfer. Clonal cell lines A2, A7 and A8 had blastocyst rates of 11.7% a , 16.7% ab and 20.0% b , respectively ( ab p < 0.05). Adult somatic cells included kidney, mammary (large), and mammary (small) also had different blastocyst rates ( ab p < 0.05) of 4.2% a , 10.7% ab and 18.3% b , respectively. For clonal donor cells and for adult somatic cell groups the donor cells with the highest blastocyst rates also had methylation profiles with the lowest similarity to the methylation profiles of the in vivo-produced blastocysts. Conversely, the donor cells with the lowest blastocyst rates had methylation profiles with the highest similarity to the methylation profiles of the in vivo-produced blastocysts. Our findings show there is an inverse correlation to the similarity of the methylation profiles of the donor cells and the in vivo-produced embryos, and to the blastocyst rates following SCNT. ABBREVIATIoNSA2, A7, A8, designation for three different donor cell populations; ANOVA, analysis of variance; BSA, bovine serum albumin;

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Mice cloned from embryonic stem cells

Cited by 436 publications

References 37 publications

Mouse cloning and somatic cell reprogramming using electrofused blastomeres

Mouse cloning and somatic cell reprogramming using electrofused blastomeres

Propagation of an infertile hermaphrodite mouse lacking germ cells by using nuclear transfer and embryonic stem cell technology

Correlation of Developmental Differences of Nuclear Transfer Embryos Cells to the Methylation Profiles of Nuclear Transfer Donor Cells in Swine

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