Production of Live Calves Derived from Embryonic Stem-Like Cells Aggregated with Tetraploid Embryos1

Campbell, Keith; Galli, Cesare; Akiyama, Kiyoshi; Iwasaki, Setsuo

doi:10.1095/biolreprod62.2.470

Cited by 88 publications

(60 citation statements)

References 25 publications

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“…37 These methodologies have been used in cattle, and chimera animals were generated. 38,39 However, the establishment of pluripotent ESCs from bovine embryos has proven unsuccessful, and the generation of transgenic bovine chimeric offspring was only possible using nuclear transplantation, where transgenic embryonic stem (ES)-like cells were produced from modified fetal bovine fibroblasts. 40 Transgenic porcine chimeras were generated using primordial germ cell (PGCs)-derived colonies, 41 and PGCs can become EGCs by in vitro or in vivo differentiation.…”

Section: Strategies For the Generation Of Transgenic Cattlechallengesmentioning

confidence: 99%

Transgenic bovine as bioreactors: Challenges and perspectives

et al. 2016

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The use of recombinant proteins has increased in diverse commercial sectors. Various systems for protein production have been used for the optimization of production and functional protein expression. The mammary gland is considered to be a very interesting system for the production of recombinant proteins due to its high level of expression and its ability to perform post-translational modifications. Cows produce large quantities of milk over a long period of lactation, and therefore this species is an important candidate for recombinant protein expression in milk. However, transgenic cows are more difficult to generate due to the inefficiency of transgenic methodologies, the long periods for transgene detection, recombinant protein expression and the fact that only a single calf is obtained at the end of each pregnancy. An increase in efficiency for transgenic methodologies for cattle is a big challenge to overcome. Promising methodologies have been proposed that can help to overcome this obstacle, enabling the use of transgenic cattle as bioreactors for protein production in milk for industry.

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Section: Strategies For the Generation Of Transgenic Cattlechallengesmentioning

confidence: 99%

Transgenic bovine as bioreactors: Challenges and perspectives

et al. 2016

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“…In these chimeras, the tetraploid cells can contribute to trophoblast derivatives, but not the embryos/fetus proper (Cross 2001). Tetraploid chimeras in both mice (Eggan et al 2001) and cattle (Iwasaki et al 2000) do better than regular clones, implying that trophoblast lineage defects in clones account for at least some of the pregnancy complications.…”

Section: Placental Mal-development In Cloned Animalsmentioning

confidence: 99%

Placental function in development and disease

Cross

2006

Reprod. Fertil. Dev.

112

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Abstract. The placenta is an organ that clinicians and embryologists would all agree is important for pregnancy success. Unfortunately, however, they too often ignore it when they are exploring causes for embryonic, fetal and perinatal complications. The core function of the placenta is to mediate the transport of nutrients between the maternal and fetal circulation, but it also has critical endocrine functions that alter different maternal physiological systems in order to sustain pregnancy. Both its development and ongoing functions can be dynamically regulated by environmental factors, including nutrient status and tissue oxygenation. In recent years, mainstream attention has begun to shift onto the placenta and it is now becoming clear that placental pathology is associated with several complications in human and animal pregnancies, including embryonic lethality, fetal growth restriction, pre-eclampsia and the high rates of fetal deaths observed after nuclear transfer (cloning).

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“…However, developmental abnormalities can arise during pregnancy and/or postnatal, limiting the widespread application of this technology [1][2][3]. Tetraploid embryos are used in the production of transgenic mice as tetraploid complementation with mouse embryonic stem (ES) cells increases the success rate of cloning [4][5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

“…The third method is to fuse two diploid blastomeres at the 2-cell stage embryo with either polyethylene glycol (PEG) [18,19], inactivated Sendai virus [20] or by electrofusion. Among these fusion methods, electrofusion is the most widely used tool for the production of tetraploid embryos [1,21,22] as it is safer than the chemical-or virus-mediated methods and more convenient and economical than microsurgical injection [14].…”

Section: Introductionmentioning

confidence: 99%

Efficient Production of Bovine Tetraploid Embryos by Electrofusion of In Vitro Produced Two-Cell Stage Embryos

Wang¹,

Song²

2016

JFIV Reprod Med Genet

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Tetraploid complementation has been used to improve the production of cloned animals. The main objective of this study was to improve the efficiency of bovine tetraploid embryo production and the development potential of bovine tetraploid embryos into blastocysts. We assessed early embryonic cleavage timing and established the defined time quantum to collect synchronous 2-cell stage bovine embryos for electrofusion. Different electrofusion protocols were also tested. The second aim was to monitor ploidy transition by fluorescence visualization to shed lights on the nuclear fusion process of the cytoplasmic membrane-fused 2-cell stage bovine embryos. Karyotypes of day 8 blastocysts (day 0: day of electrofusion) were determined by karyotyping analysis. We found that electrofusion of in vitro produced bovine 2-cell stage embryos results in both tetraploid and diploid embryos and that blastocyst formation rates from fused 2-cell stage embryos are affected by the number of electrofusion pulses and the timing of how soon the 2-cell stage embryos are formed post insemination. We identified two distinct nuclear configurations after electrofusion of 2-cell stage embryos, each of which is uniquely related to the formation of tetraploid or diploid embryos. Based on the observation that tetraploid and diploid embryos derived from fused 2-cell stage embryos undergo different timings to become 2-cell stage embryos again, diploid and tetraploid 2-cell embryos can be readily separated after electrofusion. Finally, our study established an experimental protocol for the effective production of bovine tetraploid embryos by electrofusion of 2-cell embryos.

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Production of Live Calves Derived from Embryonic Stem-Like Cells Aggregated with Tetraploid Embryos1

Cited by 88 publications

References 25 publications

Transgenic bovine as bioreactors: Challenges and perspectives

Transgenic bovine as bioreactors: Challenges and perspectives

Placental function in development and disease

Efficient Production of Bovine Tetraploid Embryos by Electrofusion of In Vitro Produced Two-Cell Stage Embryos

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