scholarly journals Interspecies Nuclear Transfer Embryos Reconstructed from Cat Somatic Cells and Bovine Ooplasm

2008 ◽  
Vol 54 (2) ◽  
pp. 142-147 ◽  
Author(s):  
Ampika THONGPHAKDEE ◽  
Shuji KOBAYASHI ◽  
Kei IMAI ◽  
Yasushi INABA ◽  
Mariko TASAI ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells ◽  
Interspecies Nuclear Transfer

scholarly journals 57EMBRYO DEVELOPMENT FOLLOWING INTERSPECIES NUCLEAR TRANSFER OF AFRICAN BUFFALO (SYNCERUS CAFFER), BONTEBOK (DAMALISCUS DORCUS DORCUS) AND ELAND (TAUROTRAGUS ORYX) SOMATIC CELLS INTO BOVINE CYTOPLASTS

2004 ◽  
Vol 16 (2) ◽  
pp. 150 ◽  
Author(s):  
M. Matshikiza ◽  
P. Bartels ◽  
G. Vajta ◽  
F. Olivier ◽  
T. Spies ◽  
...  
Keyword(s):  
South Africa ◽  
Nuclear Transfer ◽  
Somatic Cells ◽  
Blastocyst Stage ◽  
Critically Endangered ◽  
African Buffalo ◽  
Syncerus Caffer ◽  
Cell Stage ◽  
Wildlife Species ◽  
Interspecies Nuclear Transfer

Wildlife conservation requires traditional as well as innovative conservation strategies in order to preserve gene and species diversity. Interspecies nuclear transfer has the potential to conserve genes from critically endangered wildlife species where few or no oocytes are available from the endangered species, and where representative cell lines have been established for the wildlife population while numbers were still abundant. The purpose of this study was to investigate the developmental ability of embryos reconstructed with transfer of somatic cells from the African buffalo (Syncerus caffer), bontebok (Damaliscus dorcus dorcus) and eland (Taurotragus oryx) to enucleated domestic cattle (Bos taurus) oocytes. Skin tissue from the three wildlife species were collected by surgically removing approx. 1.0×1.0cm ear skin notches from animals immobilized with a combination of etorphine hydrochloride (M99; South Africa) and azaperone (Stressnil, South Africa). The biopsies were placed into physiological saline and transported to the laboratory at 4°C within 2h, cleaned with chlorohexidine gluconate and sliced finely in Minimal Essential Medium supplemented with 10% fetal calf serum. The resultant tissue explants were treated as previously described (Baumgarten and Harley 1995 Comp. Biochem. Physiol. 110B, 37–46) and actively growing fibroblast cultures made available for the nuclear transfer process. Nuclear transfer was performed using the HMC technique (Vajta et al., 2003 Biol. Reprod. 68, 571–578) using slaughterhouse-derived bovine oocytes. Culture was performed in SOFaaci (Vajta et al., 2003 Biol. Reprod. 68, 571–578) medium supplemented with 5% cattle serum using WOWs (Vajta et al., Mol. Reprod. Dev. 50, 185–191). Two identical replicates were made with somatic cells of each species. After successful reconstruction, 57, 42 and 48 nuclear transferred and activated buffalo, bontebok and eland embryos were cultured, respectively. All except for 2 buffalo embryos cleaved; 22 (39%) developed to or over the 8-cell stage, and 2 (3.5%) of them to the blastocyst stage. All but 3 bontebok embryos cleaved, 17 (40%) developed to or over the 8-cell stage, but none of them reached the compacted morula or blastocyst stage. Sixteen (33%) of the eland embryos developed to or over the 8-cell stage with one (2%) reaching the blastocyst stage. In conclusion, buffalo, bontebok and eland embryos developed from reconstruction using their respective somatic cells combined with bovine cytoplasts, however, in vitro developmental ability to the blastocyst stage was limited. Additional basic research that establishes the regulative mechanisms involved with early preimplantation development together with optimising nuclear transfer techniques may have the potential to one day play a role in the conservation of critically endangered wildlife species.

IN VITRO DEVELOPMENT OF INTERSPECIES NUCLEAR TRANSFER USING PORCINE OOCYTES WITH GOAT AND MOUSE SOMATIC CELLS

2007 ◽  
Vol 77 (Suppl_1) ◽  
pp. 237-237
Author(s):  
Yanshi Quan ◽  
Kenji Naruse ◽  
Baek Chul Kim ◽  
Su Min Choi ◽  
Rong Xun Han ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells ◽  
In Vitro Development ◽  
Interspecies Nuclear Transfer ◽  
Vitro Development

Cloned blastocysts produced by nuclear transfer from somatic cells in cynomolgus monkeys (Macaca fascicularis)

Primates ◽  
2007 ◽  
Vol 48 (3) ◽  
pp. 232-240 ◽  
Author(s):  
Junko Okahara-Narita ◽  
Hideaki Tsuchiya ◽  
Tatsuyuki Takada ◽  
Ryuzo Torii
Keyword(s):  
Nuclear Transfer ◽  
Macaca Fascicularis ◽  
Somatic Cells ◽  
Cynomolgus Monkeys

Cloned rabbits produced by nuclear transfer from adult somatic cells

2002 ◽  
Vol 20 (4) ◽  
pp. 366-369 ◽  
Author(s):  
Patrick Chesné ◽  
Pierre G. Adenot ◽  
Céline Viglietta ◽  
Michel Baratte ◽  
Laurent Boulanger ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells

Nuclear transplantation and the control of gene activity in animal development

1970 ◽  
Vol 176 (1044) ◽  
pp. 303-314 ◽  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells ◽  
Gene Activity ◽  
Protein Fractions ◽  
Nuclear Transplantation ◽  
Animal Development ◽  
Cytoplasmic Proteins ◽  
Intracellular Movement ◽  
Selective Accumulation ◽  
Structure And Metabolism

The transplantation of nuclei from differentiated or determined somatic cells to enucleated frogs’ eggs consistently leads to a complete and clearly recognizable change of gene activity. Within 1 to 2 h of nuclear transfer, somatic nuclei have come to resemble in structure and metabolism the zygote nuclei of fertilized eggs. The change in gene activity therefore takes place very soon after nuclear transfer and results from an effect of egg cytoplasm. The induced change in gene activity is associated with a selective accumulation of cytoplasmic proteins in transplanted nuclei. Examples are given of various ways in which nuclear transplantation and microinjection can be used to elucidate the intracellular movement of proteins and the effect of known protein fractions on gene activity.

scholarly journals Erratum: Production of gene-targeted sheep by nuclear transfer from cultured somatic cells

Nature ◽  
2000 ◽  
Vol 408 (6808) ◽  
pp. 120-120 ◽  
Author(s):  
K. J. McCreath ◽  
J. Howcroft ◽  
K. H. S. Campbell ◽  
A. Colman ◽  
A. E. Schnieke ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Somatic Cells

scholarly journals Differential In Vivo Binding Dynamics of Somatic and Oocyte-specific Linker Histones in Oocytes and During ES Cell Nuclear Transfer

2005 ◽  
Vol 16 (8) ◽  
pp. 3887-3895 ◽  
Author(s):  
Matthias Becker ◽  
Antje Becker ◽  
Faiçal Miyara ◽  
Zhiming Han ◽  
Maki Kihara ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Somatic Cells ◽  
Linker Histone ◽  
Binding Properties ◽  
Paternal Genome ◽  
Linker Histones ◽  
In Vivo Binding

The embryonic genome is formed by fusion of a maternal and a paternal genome. To accommodate the resulting diploid genome in the fertilized oocyte dramatic global genome reorganizations must occur. The higher order structure of chromatin in vivo is critically dependent on architectural chromatin proteins, with the family of linker histone proteins among the most critical structural determinants. Although somatic cells contain numerous linker histone variants, only one, H1FOO, is present in mouse oocytes. Upon fertilization H1FOO rapidly populates the introduced paternal genome and replaces sperm-specific histone-like proteins. The same dynamic replacement occurs upon introduction of a nucleus during somatic cell nuclear transfer. To understand the molecular basis of this dynamic histone replacement process, we compared the localization and binding dynamics of somatic H1 and oocyte-specific H1FOO and identified the molecular determinants of binding to either oocyte or somatic chromatin in living cells. We find that although both histones associate readily with chromatin in nuclei of somatic cells, only H1FOO is capable of correct chromatin association in the germinal vesicle stage oocyte nuclei. This specificity is generated by the N-terminal and globular domains of H1FOO. Measurement of in vivo binding properties of the H1 variants suggest that H1FOO binds chromatin more tightly than somatic linker histones. We provide evidence that both the binding properties of linker histones as well as additional, active processes contribute to the replacement of somatic histones with H1FOO during nuclear transfer. These results provide the first mechanistic insights into the crucial step of linker histone replacement as it occurs during fertilization and somatic cell nuclear transfer.

52 INITIATION OF PREGNANCIES IN SOUTH AFRICAN RIVERINE RABBIT (BUNOLAGUS MONTICULARES) BY INTERSPECIES NUCLEAR TRANSFER USING ADIPOSE-DERIVED SOMATIC CELLS

2008 ◽  
Vol 20 (1) ◽  
pp. 106
Author(s):  
M. J. Sansinena ◽  
D. Owiny ◽  
R. S. Denniston ◽  
D. Salamone ◽  
D. Barry
Keyword(s):  
In Vitro Culture ◽  
Nuclear Transfer ◽  
Uv Light ◽  
Uv Exposure ◽  
Domestic Rabbit ◽  
Donor Cells ◽  
Significant Difference ◽  
Preliminary Study ◽  
Interspecies Nuclear Transfer

The riverine rabbit (Bunolagus monticulares), one of South Africa's most threatened mammals, with an estimated population size under 250, was upgraded from endangered to critically endangered in 2002. The low number of riverine rabbits precludes any attempts of nuclear transfer (NT) using intraspecific oocytes; therefore, the overall aim of this study was to assess the ability of the domestic rabbit (Oryctolagus cuniculus) oocyte to reprogram the somatic cell of the endangered riverine rabbit by interspecies NT. A preliminary study evaluated the effect of timing of enucleation after induction of ovulation (h post-hCG). A second study assessed the effects of two activation protocols. In addition, since the unique characteristics of the rabbit zona pellucida affect the speed of micromanipulation, different exposure periods to UV light at enucleation were evaluated. Adult domestic Californian rabbits were treated with eCG for 72 h, and ovulation was induced by hCG administration. Oocytes were collected by retrograde flushing at 12–14 h or 16–18 h post-hCG administration and stripped of cumulus investments with 0.5% hyaluronidase in Ca-Mg-free PBS. Metaphase-II oocytes were selected by visualizing the first polar body. Oocytes were stained with 2 mg mL–1 Hoechst 33342 for 5 min, and metaphase plates were removed with a 25–30 μm (O.D.) borosilicate beveled, spiked pipette after exposure to <5 or 30–40 s of UV light. Adult adipose-derived riverine rabbit fibroblasts grown to confluency in DMEM with 10% FCS were used as donor cells and fused with 2 consecutive DC pulses (3.2 kV cm–1, 45 μs). After reconstruction, couplets were randomly assigned for activation by either a second set of electrical pulses or incubation with ionomycin, followed by 1 h of incubation in 2 mm 6-DMAP. Embryos were co-cultured with a bovine oviductal cell monolayer in DMEM with 10% FCS and assessed for cleavage after 36 h of in vitro culture. There was a significant difference in the number of cleaved embryos from oocytes collected at 12–14 h post-hCG (n = 50) or 16–18 h post-hCG (n = 51) administration (57% v. 0% cleaved; P < 0.05). No significant difference was detected in embryos developing after electrofusion v. ionomycin activation treatments. However, a significantly greater number (P < 0.05) of embryos cleaved from oocytes exposed to <5 s UV than from oocytes exposed to 30–40 s UV (Table 1). A total of 20 embryos (4-cell to 16-cell stages) were surgically transferred to the oviducts of 4 adult New Zealand white synchronized recipients after 48 h of in vitro culture. Two recipients (<5 s UV exposure treatment group) were diagnosed pregnant by abdominal palpation at 15 days post-transfer; pregnancies were subsequently lost by Day 30, with placental tissues recovered. This preliminary study indicates the domestic rabbit oocyte is capable of reprogramming riverine rabbit donor cells. In addition, the time of oocyte collection after ovulation induction and the UV exposure period during enucleation have an effect on the efficiency of interspecies NT and embryo development in this species. Table 1. Effect of UV exposure during enucleation on the in vitro development of interspecies nuclear transfer riverine rabbit embryos

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