Intergeneric somatic cell nucleus transfer in marbled cat and flat-headed cat

Thongphakdee, Ampika; Siriaroonrat, Boripat; Manee-in, Sukanya; Klincumhom, Nuttha; Kamolnorranath, Sumate; Chatdarong, Kaywalee; Techakumphu, Mongkol

doi:10.1016/j.theriogenology.2009.09.001

Cited by 23 publications

(11 citation statements)

References 36 publications

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“…The successful application of iSCNT in felids was demonstrated by the birth of African wild cats (Felis silvestris lybica) (Gó mez et al 2004b) and sand cats (Felis margarita) (Gó mez et al 2008), as well as by pregnancies reported from embryos generated by the fusion of leopard cat (Prionailurus bengalensis) cells with Dc enucleated oocytes (Yin et al 2006). Despite these achievements, pregnancies and births after iSCNT are still elusive, as shown in several reports (Thongphakdee et al 2010, Gó mez et al 2011, Imsoonthornruksa et al 2012.…”

Section: Introductionmentioning

confidence: 99%

Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression

Moro¹,

Hiriart²,

Buemo³

et al. 2015

Reproduction

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The aim of this study was to evaluate the capacity of domestic cat (Dc, Felis silvestris) oocytes to reprogram the nucleus of cheetah (Ch, Acinonyx jubatus) cells by interspecies SCNT (iSCNT), by using embryo aggregation. Dc oocytes were in vitro matured and subjected to zona pellucida free (ZP-free) SCNT or iSCNT, depending on whether the nucleus donor cell was of Dc or Ch respectively. ZP-free reconstructed embryos were then cultured in microwells individually (Dc1X and Ch1X groups) or in couples (Dc2X and Ch2X groups). Embryo aggregation improved in vitro development obtaining 27.4, 47.7, 16.7 and 28.3% of blastocyst rates in the Dc1X, Dc2X, Ch1X and Ch2X groups, respectively (P!0.05). Moreover, aggregation improved the morphological quality of blastocysts from the Dc2X over the Dc1X group. Gene expression analysis revealed that Ch1X and Ch2X blastocysts had significantly lower relative expression of OCT4, CDX2 and NANOG than the Dc1X, Dc2X and IVF control groups. The OCT4, NANOG, SOX2 and CDX2 genes were overexpressed in Dc1X blastocysts, but the relative expression of these four genes decreased in the Dc2X, reaching similar relative levels to those of Dc IVF blastocysts. In conclusion, Ch blastocysts were produced using Dc oocytes, but with lower relative expression of pluripotent and trophoblastic genes, indicating that nuclear reprogramming could be still incomplete. Despite this, embryo aggregation improved the development of Ch and Dc embryos, and normalized Dc gene expression, which suggests that this strategy could improve full-term developmental efficiency of cat and feline iSCNT embryos.Reproduction (2015) 150 1-10

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Section: Introductionmentioning

confidence: 99%

Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression

Moro¹,

Hiriart²,

Buemo³

et al. 2015

Reproduction

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“…2006), the marbled cat (Pardofelis marmorata) (Thongphakdee et al. 2006) and the flat‐headed cat (Prionailurus planiceps) (Thongphakdee et al. 2010).…”

Section: Introductionmentioning

confidence: 99%

Cell Cycle Synchronization of Skin Fibroblast Cells in Four Species of Family Felidae

Wittayarat

Thongphakdee

Saikhun

et al. 2012

Reprod Domestic Animals

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This study was examined whether the species of felid affects synchronization accuracy at the G0/G1 stage of the cell cycle and the occurrence of apoptosis by different protocols, such as serum starvation, confluent and roscovitine treatment. Skin fibroblast cells were obtained from the Asian golden cat, marbled cat, leopard and Siamese cat. The cells from each animal were treated with either serum starvation for 1-5 days, cell confluency-contact inhibition for 5 days or roscovitine at various concentrations (7.5-30 μm). Flow cytometric analysis revealed that serum starvation for 3 days provided the highest cell population arrested at the G0/G1 stage, irrespective of the felid species. In all species, 100% confluency gave a significantly higher percentage of cells arrested at the G0/G1 stage compared with the non-treated control cells. The effects of roscovitine treatment and the appropriate concentration on the rates of G0/G1 cells differed among the felid species. Serum starvation for more than 4 days in the marbled cat and Siamese cat and roscovitine treatment with 30 μm in the Asian golden cat and leopard increased the rates of apoptosis. In conclusion, different felid species responded to different methods of cell cycle synchronization. Asian golden cat and Siamese cat fibroblast cells were successfully synchronized to G0/G1 stage using the serum starvation and roscovitine treatment, whereas only confluency-contact inhibition treatment induced cell synchronization in the leopard. Moreover, these three methods did not successfully induce cell synchronization of the marbled cat. These findings may be valuable for preparing their donor cells for somatic cell nuclear transfer in the future.

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“…Specifically on felids, the successful application of iSCNT was confirmed after birth of African wild cats (Felis silvestris lybica; G omez et al 2004) and sand cats (Felis margarita;G omez et al 2008), and pregnancies obtained in leopard cats (Prionailurus bengalensis, Yin et al 2006). Despite these achievements, it is still very difficult to obtain pregnancies and births after iSCNT, as was shown in several reports (Thongphakdee et al 2010;G omez et al 2011;Imsoonthornruksa et al 2012). The causes of its inefficiency are many, and they could be related to failures in nuclear-cytoplasmic communications (St John et al 2004) and to the basis of nuclear transfer itself (Niemann et al 2008;Palmieri et al 2008).…”

Section: Introductionmentioning

confidence: 93%

“…Despite these achievements, it is still very difficult to obtain pregnancies and births after iSCNT, as was shown in several reports (Thongphakdee et al. ; Gómez et al. ; Imsoonthornruksa et al.…”

Section: Introductionmentioning

confidence: 99%

Tiger, Bengal and Domestic Cat Embryos Produced by Homospecific and Interspecific Zona‐Free Nuclear Transfer

Moro

Jarazo

Buemo

et al. 2015

Reprod Domestic Animals

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The aim of this study was to evaluate three different cloning strategies in the domestic cat (Felis silvestris) and to use the most efficient to generate wild felid embryos by interspecific cloning (iSCNT) using Bengal (a hybrid formed by the cross of Felis silvestris and Prionailurus bengalensis) and tiger (Panthera tigris) donor cells. In experiment 1, zona-free (ZP-free) cloning resulted in higher fusion and expanded blastocyst rates with respect to zona included cloning techniques that involved fusion or injection of the donor cell. In experiment 2, ZP-free iSCNT and embryo aggregation (2X) were assessed. Division velocity and blastocyst rates were increased by embryo aggregation in the three species. Despite fewer tiger embryos than Bengal and cat embryos reached the blastocyst stage, Tiger 2X group increased the percentage of blastocysts with respect to Tiger 1X group (3.2% vs 12.1%, respectively). Moreover, blastocyst cell number was almost duplicated in aggregated embryos with respect to non-aggregated ones within Bengal and tiger groups (278.3 ± 61.9 vs 516.8 ± 103.6 for Bengal 1X and Bengal 2X groups, respectively; 41 vs 220 ± 60 for Tiger 1X and Tiger 2X groups, respectively). OCT4 analysis also revealed that tiger blastocysts had higher proportion of OCT4-positive cells with respect to Bengal blastocysts and cat intracytoplasmic sperm injection blastocysts. In conclusion, ZP-free cloning has improved the quality of cat embryos with respect to the other cloning techniques evaluated and was successfully applied in iSCNT complemented with embryo aggregation.

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Intergeneric somatic cell nucleus transfer in marbled cat and flat-headed cat

Cited by 23 publications

References 36 publications

Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression

Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression

Cell Cycle Synchronization of Skin Fibroblast Cells in Four Species of Family Felidae

Tiger, Bengal and Domestic Cat Embryos Produced by Homospecific and Interspecific Zona‐Free Nuclear Transfer

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