Evaluation of the embryonic preimplantation potential of human adult somatic cells via an embryo interspecies bioassay using bovine oocytes

Illmensee, Karl; Levanduski, Mike; Zavos, Panayiotis M.

doi:10.1016/j.fertnstert.2005.10.026

Cited by 42 publications

(20 citation statements)

References 42 publications

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“…Our results were not different from several other recent reports (Chung et al 2009, Lagutina et al 2010, who had also reported that iSCNT embryos failed to reactivate the embryonic genome and arrested at the stage of EGA specific to the recipient oocyte. Conversely, inter-genus (Oh et al 2006, Yin et al 2006, inter-family (Dominko et al 1999, Zhao et al 2007, inter-order (Yang et al 2003, Wen et al 2005, Illmensee et al 2006, and even inter-class iSCNT (Chen et al 2002, Liu et al 2004 nuclear transfer embryos were reported to have Pig-mouse cytoplasmic hybrid embryos developed to the blastocyst stage. However, these later reports also observed that the majority of iSCNT embryos arrested around the stage of EGA and only a very small percent of embryos developed to blastocyst stage.…”

Section: Discussionmentioning

confidence: 99%

Nuclear–cytoplasmic incompatibility and inefficient development of pig–mouse cytoplasmic hybrid embryos

Amarnath

Choi²,

Moawad³

et al. 2011

Reproduction

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Inter-species somatic cell nuclear transfer (iSCNT) embryos usually fail to develop to the blastocyst stage and beyond due to incomplete reprogramming of donor cell. We evaluated whether using a karyoplast that would require less extensive reprogramming such as an embryonic blastomere or the meiotic spindle from metaphase II oocytes would provide additional insight into the development of iSCNT embryos. Our results showed that karyoplasts of embryonic or oocyte origin are no different from somatic cells; all iSCNT embryos, irrespective of karyoplast origin, were arrested during early development. We hypothesized that nuclear-cytoplasmic incompatibility could be another reason for failure of embryonic development from iSCNT. We used pig-mouse cytoplasmic hybrids as a model to address nuclear-cytoplasmic incompatibility in iSCNT embryos. Fertilized murine zygotes were reconstructed by fusing with porcine cytoplasts of varying cytoplasmic volumes (1/10 (small) and 1/5 (large) total volume of mouse zygote). The presence of pig cytoplasm significantly reduced the development of mouse zygotes to the blastocyst stage compared with control embryos at 120 h post-human chorionic gondotropin (41 vs 6 vs 94%, P!0.05; 1/10, 1/5, control respectively). While mitochondrial DNA copy numbers remained relatively unchanged, expression of several important genes namely Tfam, Polg, Polg2, Mfn2, Slc2a3 (Glut3), Slc2a1 (Glut1), Bcl2, Hspb1, Pou5f1 (Oct4), Nanog, Cdx2, Gata3, Tcfap2c, mt-Cox1 and mt-Cox2 was significantly reduced in cytoplasmic hybrids compared with control embryos. These results demonstrate that the presence of even a small amount of porcine cytoplasm is detrimental to murine embryo development and suggest that a range of factors are likely to contribute to the failure of inter-species nuclear transfer embryos.

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Section: Discussionmentioning

confidence: 99%

Nuclear–cytoplasmic incompatibility and inefficient development of pig–mouse cytoplasmic hybrid embryos

Amarnath

Choi²,

Moawad³

et al. 2011

Reproduction

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“…Llama and dromedary belong to same family Camelidae, have similar reproductive physiology but their size varies greatly. Even though many studies have reported production of iSCNT morulae and blastocysts when nucleus donor cells and recipient oocytes had a very distant taxonomical relation, like interfamily bovine—pig [6,30], interorder cat—and panda—rabbit [31], camel—and Tibetan antelope—rabbit [32], human—rabbit [33], dog—pig [34], tiger—pig [35], human—bovine [36,37], human—ovine [38], or interclass chicken—rabbit [39] combinations. We did not transfer theses embryos in the present study due to lack of sufficient recipients.…”

Section: Discussionmentioning

confidence: 99%

First cloned Bactrian camel (Camelus bactrianus) calf produced by interspecies somatic cell nuclear transfer: A step towards preserving the critically endangered wild Bactrian camels

2017

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Studies were conducted to explore the possibility of employing dromedary camel (Camelus dromedarius) oocytes as recipient cytoplasts for the development of interspecies somatic cell nuclear transfer (iSCNT) embryos using skin fibroblast cells of an adult Bactrian camel (Camelus bactrianus) and Llama (Llama glama) as donor nuclei. Also, the embryos reconstructed with Bactrian cells were transferred into the uterus of synchronized dromedary camel recipients to explore the possibility of using them as surrogate mothers. Serum-starved skin fibroblast cells were injected into the perivitelline space of enucleated mature oocytes, collected from super-stimulated dromedary camels, and fused using an Eppendorf electroporator. After activation with 5μM ionomycin and 6-dimethylaminopurine, they were cultured at 38.5°C in an atmosphere of 5% CO2, 5% O2, and 90% N2 in air. In experiment 1, Day 7 blastocysts were stained with Hoechst to count their cell numbers, while in experiment 2, they were transferred to synchronized dromedary recipients. A lower number (P < 0.05) of blastocysts were obtained from reconstructs utilizing fibroblast cells from Llama when compared with those reconstructed with dromedary and Bactrian fibroblast cells. However, no difference was observed in their cell numbers. In experiment 2, a higher (P < 0.05) proportion of blastocysts were obtained from the cleaved embryos reconstructed with Bactrian fibroblast cells when compared to those reconstructed with dromedary cells. Twenty-six Day 7 blastocysts reconstructed with Bactrian cells were transferred to 23 synchronized dromedary recipients with 5 pregnancies established on Day 30, however, only one of the pregnancies developed to term and a healthy calf weighing 33 kgs was born after completing 392 days of gestation. Unfortunately, the calf died on day 7 due to acute septicemia. In conclusion, the present study reports, for the first time, birth of a cloned Bactrian calf by iSCNT using dromedary camel as a source for oocytes as well as a surrogate for carrying the pregnancy to term.

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“…There has been at least one report of successful hES cell line derivation following SCNT of adult human fibroblasts into enucleated rabbit oocytes,15 and another more controversial into bovine oocytes 16. Although the status of the rabbit/human mitochondria was not addressed in Chen’s paper,15 the expectation is that the transfer of an intact human cell into an enucleated rabbit oocyte would result in the rabbit proteins and mitochondria being replaced with human versions over time 17.…”

Section: What We Are Speaking Ofmentioning

confidence: 99%

Fearing a non-existing Minotaur? The ethical challenges of research on cytoplasmic hybrid embryos

Camporesi¹,

Boniolo

2008

J Med Ethics

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Evaluation of the embryonic preimplantation potential of human adult somatic cells via an embryo interspecies bioassay using bovine oocytes

Cited by 42 publications

References 42 publications

Nuclear–cytoplasmic incompatibility and inefficient development of pig–mouse cytoplasmic hybrid embryos

Nuclear–cytoplasmic incompatibility and inefficient development of pig–mouse cytoplasmic hybrid embryos

First cloned Bactrian camel (Camelus bactrianus) calf produced by interspecies somatic cell nuclear transfer: A step towards preserving the critically endangered wild Bactrian camels

Fearing a non-existing Minotaur? The ethical challenges of research on cytoplasmic hybrid embryos

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