Efficient Production of Cloned Bovine Embryos Using cdc2 kinase Inhibitor

Yoo, Jae Gyu; Sy, Choe; Gj, Rho

doi:10.1046/j.0936-6768.2003.00461.x

Cited by 12 publications

(8 citation statements)

References 62 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This result is similar to the previous studies on DMAP treatment for oocytes activation in cattlem, which displayed the alterations of karyokinesis during the first cell cycle, causing the failure of the second PB extrusion and re-entering of some nuclei to S-phase of the cell cycle without having passed through metaphase (De La Fuente and King, 1998). Slimane-Bureau and King (2002) and Yoo et al (2003) also opined that cattle NT embryos with use of DMAP are associated with high levels of embryonic mortality and often with congenital malformation by chromosomal abnormalities.…”

Section: Discussionsupporting

confidence: 90%

Development of cloned pig embryos by nuclear transfer following different activation treatments

Kim¹,

Lee²,

Ock³

et al. 2004

Molecular Reproduction Devel

View full text Add to dashboard Cite

This study compared the effects of activation treatments on the development and ploidy of nuclear transferred (NT) pig embryos. After in vitro maturation of oocytes collected from the slaughterhouse, oocytes were enucleated and reconstructed by transfer of donor cells and fusion with three DC pulses (1.4 kV/cm, 20 musec). Oocytes were pulsed thrice electrically with 1.4 kV/cm for 20 musec and NT eggs were divided into three treatment groups: Group 1 (no further treatment), Group 2 (10 mug/mL cycloheximide [CHX], 3 hr), and Group 3 (1.9 mM 6-dimethylaminopurine [DMAP], 3 hr). All the eggs were cultured in sets of 30 in 60 muL drops of NCSU-23 supplemented with 4 mg/mL fatty acid free BSA, and compared for the rates of development and ploidy. The rates of cleavage, development, and total cell number of parthenotes in Group 3 were significantly (P < 0.05) higher than those in Groups 1 and 2. Cleavage rates of NT embryos in Group 1 were significantly (P < 0.05) lower than those in Groups 2 and 3 (73% vs. 81% and 82%, respectively). Development into blastocyst stage and total cell number of NT embryos in Group 3 were significantly (P < 0.05) higher than those in Groups 1 and 2. Although the embryos in Group 3 had higher development, approximately 58% of NT embryos evaluated were abnormal ploidy (6% haploidy, 9% polyploidy, and 42% mixoploid). The results suggested that although DMAP enhanced development and higher cell number, due attention should be paid to abnormal ploidy in pig NT embryos.

show abstract

Section: Discussionsupporting

confidence: 90%

Development of cloned pig embryos by nuclear transfer following different activation treatments

Kim¹,

Lee²,

Ock³

et al. 2004

Molecular Reproduction Devel

View full text Add to dashboard Cite

show abstract

“…In addition to efficient embryo development and transgene expression, it is possible that the problems associated with cloning and ICSI‐mgt embryos could be avoided. In particular, the chemical assistance required for embryonic activation in these techniques generates chromatin defects (De La Fuente and King 1998; Yoo et al. 2003; Bhak et al.…”

Section: Discussionmentioning

confidence: 99%

Efficient Transgene Expression in IVF and Parthenogenetic Bovine Embryos by Intracytoplasmic Injection of DNA–Liposome Complexes

Vichera

Moro

Salamone

2011

Reprod Domestic Animals

View full text Add to dashboard Cite

Transgenic animals constitute an important tool with many biotechnological applications. Although there have been advances in this field, we propose a novel method that may greatly increase the efficiency of transgenic animal production and thereby its application. This new technique consists of intracytoplasmic injection of liposomes, in bovine oocytes and zygotes, to introduce exogenous DNA. In the first experiment, we evaluated embryo development and EGFP expression in In Vitro Fertilization (IVF) embryos injected with different concentrations of exogenous DNA-liposome complexes (0.5, 5, 50, 500 ng pCX-EGFP/μl). The highest EGFP-embryos rates were obtained using 500 ng pCX-EGFP/μl. In the second experiment, we evaluated embryo development and EGFP expression following the injection of DNA-liposome complexes into pre-fertilized oocytes and presumptive zygotes, 16 and 24 h post-fertilization. Approximately 70% of the cleaved embryos and 50% of the blastocysts expressed EGFP, when egfp-liposome was injected 16 h post-fertilization. The percentages of positive embryos for the 24-h post-fertilization and pre-fertilization groups were 30.1 and 6.3, respectively. Blastocysts that developed from injected zygotes were analysed by PCR, confirming the presence of transgene in all embryos. Finally, we examined the embryo development and EGFP expression of parthenogenetic embryos that resulted from the injection of egfp-liposome complexes into pre-activated oocytes, and 3 and 11 h post-activated oocytes. The group with the highest expression rate (48.4%) was the one injected 3 h post-activation. In summary, this study reports the efficient, reproducible and fast production of IVF and parthenogenetic embryos expressing EGFP, by the intracytoplasmic injection of liposomes to introduce the foreign DNA.

show abstract

“…Despite this, these treatments produced rates of diploid embryos that did not differ from Io-DMAP, whereas Io-DMAP treatment presented higher rates of polyploidy than all DhL treatments. A high incidence of polyploidy has been observed by other authors at the blastocyst stage of parthenogenetic (De La Fuente and King, 1998;Winger et al, 1997) and SCNT bovine embryos (Bhak et al, 2006;Yoo et al, 2003) after DMAP activation. Some have attributed these anomalies to the accelerated pronuclear formation and premature DNA (þ) Reconstituted embryos by enucleation and intracytoplasmic cell injection.…”

Section: Discussionmentioning

confidence: 86%

Activation with Ionomycin followed by Dehydroleucodine and Cytochalasin B for the Production of Parthenogenetic and Cloned Bovine Embryos

Canel

Bevacqua

Fernández-Martı́n

et al. 2010

Cellular Reprogramming

View full text Add to dashboard Cite

In this work, Dehydroleucodine (DhL) was evaluated as a chemical activator of bovine oocytes and somatic cell nuclear transfer (SCNT) reconstituted embryos. Oocytes were activated with 5 microM Ionomycin (Io) and exposed for 3 h to 1 or 5 microM DhL alone (Io-Dhl1 or Io-DhL5) or combined with Cytochalasin B (Io-DhL1/CB; Io-DhL5/CB). Control groups were Io (Io), Io followed by 1.9 mM 6-Dimethylaminopurine (Io-6DMAP), and embryos produced by in vitro fertilization (IVF). Pronuclear formation and development to blastocysts of activated oocytes were evaluated. Embryos obtained by the DhL concentration that induced the highest blastocyst rates (1 microM) were karyotyped. An additional treatment based in Io-DhL1 plus lengthened (6-h) exposure to CB (Io-DhL1/long CB) was included to improve the proportion of diploid blastomeres. Finally, DhL combined with CB was employed to assist cloning by intracytoplasmic injection of whole cumulus cells. Results showed that DhL induces a pronuclear formation dynamic that was more similar to IVF-produced embryos than DMAP. Development to blastocyst stage was higher after activation with 1 microM DhL than with 5 microM DhL, either for groups combined or not with CB (19.15; 21.74 vs. 6.82; 0%, respectively) (p < 0.05). Io-DhL1 and Io-DhL1/CB treatments induced blastocyst-cleaved embryo ratios not statistically different from those of Io-DMAP (35.85%) and IVF (33.33%) groups (p > 0.05). Io-DhL1/long CB induced higher diploid blastomere rates than Io-Dhl1, Io-DhL1/CB and Io-DMAP (63.8 vs. 36.8; 40 and 31.6%, respectively) (p < 0.05). Moreover, all DhL treatments resulted in polyploidy rates that were lower than Io-DMAP (5.2, 12.0, 10.6, and 31.6%, respectively) (p < 0.05). Io-DhL1/CB and Io-DhL1/long CB induced cloned embryo blastocyst rates that were not significantly different from Io-DMAP (6.1, 9.4, and 18.3%, respectively) (p < 0.05). Our results indicate that Io-DhL1/long CB protocol could be useful for SCNT programs.

show abstract

Efficient Production of Cloned Bovine Embryos Using cdc2 kinase Inhibitor

Cited by 12 publications

References 62 publications

Development of cloned pig embryos by nuclear transfer following different activation treatments

Development of cloned pig embryos by nuclear transfer following different activation treatments

Efficient Transgene Expression in IVF and Parthenogenetic Bovine Embryos by Intracytoplasmic Injection of DNA–Liposome Complexes

Activation with Ionomycin followed by Dehydroleucodine and Cytochalasin B for the Production of Parthenogenetic and Cloned Bovine Embryos

Contact Info

Product

Resources

About