Production of horse foals via direct injection of roscovitine-treated donor cells and activation by injection of sperm extract

Hinrichs, Katrin; Choi, Young-Ho; Love, Charlie C; Chung, Young Sun; Varner, D.D.

doi:10.1530/rep.1.01095

Cited by 52 publications

(49 citation statements)

References 35 publications

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“…Flow cytometry showed that roscovitine treatment could not increase the proportion of cells arrested at the G0/G1 phase in comparison with the control group. These results are in agreement with those of Hinrichs et al (2006) who reported that roscovitine treatment did not significantly affect the proportion of horse cells in the G0/G1 phase. In contrast, Gómez et al (2003) showed that fibroblast cells from skin tissues of the domestic cat could be synchronised to the G0/G1 phase by roscovitine treatment as compared to cycling cells.…”

Section: Discussionsupporting

confidence: 93%

Cell cycle analysis and interspecies nuclear transfer of cat cells treated with chemical inhibitors

Wittayarat

Fujiwara

Chatdarong

et al. 2014

Acta Veterinaria Hungarica

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This study investigated the effect of chemical inhibitors on the cell-cycle synchronisation in cat fibroblast cells and evaluated the development of interspecies embryos reconstructed from cat donor cells and enucleated bovine oocytes. Cat fibroblast cells were treated with 15 μg/mL roscovitine or 0.05 μg/mL demecolcine prior to cell cycle analysis and nuclear transfer. The percentage of cat fibroblast cells arrested at the G0/G1 phase in the roscovitine group was similar to that in the control group without any treatment. The percentage of cells arrested at the G2/M phase was significantly higher in the demecolcine group than in the control group. The fusion rate of interspecies couplets was significantly greater in the roscovitine group than in the control group. Most embryos stopped the development at the 2-or 4-cell stage, and none developed into blastocysts. Chemical inhibitor-induced donor cell cycle synchronisation did not overcome developmental arrest in interspecies cloned embryos. Key words: Bovine oocyte, cell cycle synchronisation, demecolcine, feline, roscovitineInterspecies somatic cell nuclear transfer (iSCNT) is an alternative way to explore species for which oocytes and recipients are limited; these species include endangered or exotic species (Karja et al., 2006). It has been shown that bovine, sheep and rabbit oocyte cytoplasm supports in vitro development of embryos produced by interspecies nuclear transfer of somatic cells from various unrelated mammalian species, although no pregnancy has lasted to full term after the transfer of iSCNT embryos to surrogate animals (Dominko et al., 1999;White et al., 1999;Chen et al., 2002;Murakami et al., 2005). Moreover, the ability of iSCNT embryos to develop into blastocysts decreases as the taxonomic distance between the donor and recipient species increases (Beyhan et al., 2007).

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Section: Discussionsupporting

confidence: 93%

Cell cycle analysis and interspecies nuclear transfer of cat cells treated with chemical inhibitors

Wittayarat

Fujiwara

Chatdarong

et al. 2014

Acta Veterinaria Hungarica

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“…Reported equine NT blastocyst development rates, including those from our laboratory, have ranged from 3 to 10% (Galli et al 2003, Lagutina et al 2005, Hinrichs et al 2006. In contrast, equine blastocyst development after intracytoplasmic sperm injection (ICSI) of oocytes in our laboratory is typically 25-35% (Hinrichs et al 2005, Choi et al 2006a, 2006b).…”

Section: Introductionmentioning

confidence: 86%

“…Currently, only four viable horse foals resulting from adult somatic cell NT have been reported in the literature (Galli et al 2003, Lagutina et al 2005, Hinrichs et al 2006, and three mule foals have been produced from NT using fetal cells (Woods et al 2003, Vanderwall et al 2004a. As in other species, the rate of pregnancy loss in the horse after transfer of NT embryos has been high in most reports (75-100% of established pregnancies; Galli et al 2003, Woods et al 2003, Vanderwall et al 2004b, Lagutina et al 2005.…”

Section: Introductionmentioning

confidence: 99%

“…As in other species, the rate of pregnancy loss in the horse after transfer of NT embryos has been high in most reports (75-100% of established pregnancies; Galli et al 2003, Woods et al 2003, Vanderwall et al 2004b, Lagutina et al 2005. However, when roscovitinetreated donor cells were used for equine NT, in conjunction with injection of sperm extract for activation, transfer of eight blastocysts resulted in birth of two viable foals (Hinrichs et al 2006). Roscovitine treatment of donor cells was reported to significantly increase the proportion of viable calves born after NT (Gibbons et al 2002); this treatment may work by arresting donor cells in a phase of G1 that is amenable to reprogramming by the oocyte cytoplasm.…”

Section: Introductionmentioning

confidence: 99%

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Production of cloned horse foals using roscovitine-treated donor cells and activation with sperm extract and/or ionomycin

Hinrichs

Choi²,

Varner³

et al. 2007

Reproduction

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We evaluated the effect of different activation treatments on the production of blastocysts and foals by nuclear transfer. Donor cells were prepared using roscovitine treatment, which has previously been associated with increased production of viable offspring. All activation treatments were followed by culture in 6-dimethylaminopurine (6-DMAP) for 4 h. In experiment 1, blastocyst production after activation by injection of sperm extract followed by treatment with ionomycin was significantly higher than that for activation with a serial treatment of ionomycin, 6-DMAP, and ionomycin (12.5 vs 2.8%; P!0.05) and tended to be higher than that for injection of sperm extract alone (3.4%; PZ0.07). In experiment 2, there were no significant differences in blastocyst development among treatments with ionomycin once or twice, sperm extract then ionomycin, or ionomycin then sperm extract (range 4.6-7.3%). Overall, transfer of 26 blastocysts resulted in 16 pregnancies (62%) and 9 live foals (35% of transferred embryos). Treatment with sperm extract followed by ionomycin produced a live foal rate per embryo transferred of 5/10 (50%). One foal died of pneumonia 48 h post partum and one foal died at 1 week of age after complications during induction of anesthesia; the remaining seven foals are currently 10-14 months of age.

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“…penetration of the zona or the capsule with the Piezo drill, was based on our previous work using the drill for ICSI and nuclear transfer (Choi et al 2004, Hinrichs et al 2006. It appeared to result in less damage to the embryo than did microblade dissection, as reported previously in the horse and other species (Huhtinen et al 1997, Hasler et al 2002, El-Gayar & Holtz 2005.…”

Section: Primer Set Sequencementioning

confidence: 96%

Viability of equine embryos after puncture of the capsule and biopsy for preimplantation genetic diagnosis

Choi¹,

Gustafson-Seabury²,

Velez³

et al. 2010

Reproduction

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The equine embryo possesses a capsule that is considered essential for its survival. We assessed viability after breaching the capsule of early (Day 6) and expanded (Day 7 and 8) equine blastocysts by micromanipulation. The capsule was penetrated using a Piezo drill, and trophoblast biopsy samples were obtained for genetic analysis. Pregnancy rates for Day-6 embryos, which had intact zonae pellucidae at the time of recovery, were 3/3 for those biopsied immediately after recovery and 2/3 for those biopsied after being shipped overnight under warm (w28 8C) conditions. The pregnancy rates for encapsulated Day-7 expanded blastocysts were 5/6 for those biopsied immediately and 5/6 for those biopsied after being shipped overnight warm. Two of four encapsulated Day-8 blastocysts, 790 and 1350 mm in diameter, established normal pregnancies after biopsy. Nine mares were allowed to maintain pregnancy, and they gave birth to nine normal foals. Biopsied cells from eight embryos that produced foals were subjected to whole-genome amplification. Sex was successfully determined from amplified DNA in 8/8 embryos. Identification of disease-causing mutations matched in the analyses of 6/6 samples for the sodium channel, voltage-gated, type IV, alpha subunit (SCN4A) gene and in 6/7 samples for the peptidylprolyl isomerase B (PPIB) gene, in embryo-foal pairs. Thus, the capsule of the equine embryo can be breached without impairing viability. Further work is needed to determine whether this breach is transient or permanent. These findings are relevant to the understanding of equine embryo development and to the establishment of methods for micromanipulation and embryo cryopreservation in this species.

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Production of horse foals via direct injection of roscovitine-treated donor cells and activation by injection of sperm extract

Cited by 52 publications

References 35 publications

Cell cycle analysis and interspecies nuclear transfer of cat cells treated with chemical inhibitors

Cell cycle analysis and interspecies nuclear transfer of cat cells treated with chemical inhibitors

Production of cloned horse foals using roscovitine-treated donor cells and activation with sperm extract and/or ionomycin

Viability of equine embryos after puncture of the capsule and biopsy for preimplantation genetic diagnosis

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