The effect of activation treatments on the development of reconstructed bovine oocytes

Shen, Perng-Chih; Lee, S.N.; Liu, B.T.; Chu, F.H.; Wang, C.H.; Wu, J.S.; Lin, Hsiu-Lien; Cheng, W. T. K.

doi:10.1016/j.anireprosci.2007.03.019

Cited by 24 publications

(18 citation statements)

References 42 publications

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“…(Betthauser et al 2000;Yang et al 2007;Shen et al 2008). Cytochalasin B (CB) prevents the release of the second polar body after activation of the oocytes, which can result in diploid development (Fukui et al 1992), and may also help prevent fragmentation of embryos (Collas and Robl 1990).…”

Section: Introductionmentioning

confidence: 99%

Comparison of chemical, electrical, and combined activation methods for in vitro matured porcine oocytes

Liu

Cui

et al. 2014

In Vitro Cell.Dev.Biol.-Animal

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Factors influencing porcine oocyte activation were systematically studied. This study included (1) the effect of ionomycin plus various chemical agents on activation, (2) comparison of different electrical activation parameters, (3) optimization of combined activation, and (4) evaluation of the optimized protocols. The results showed that (1) blastocyst rates of ionomycin (Ion) + 6-dimethylaminopurine (6-DMAP) (29.7 ± 1.1%), Ion + cytochalasin B (CB) + cycloheximide (CHX) (29.8 ± 1.2%), Ion + CB + 6-DMAP (30.4 ± 1.6%), and Ion + CB + CHX + 6-DMAP (30.2 ± 2.7%) were significantly higher than Ion + CHX (15.8 ± 1.5%, p < 0.05); (2) the parthenogenetic blastocyst formation of electrical activation was optimal when oocytes were activated by three direct current (DC) pulses of 1.00 kV cm(-1) for 80 μs (39.5 ± 1.1%); (3) blastocyst rates of DC + CB + CHX (55.4 ± 1.2%) and DC + CB + 6-DMAP (50.4 ± 2.9%) were significantly higher than DC + 6-DMAP, DC + CB + CHX + 6-DMAP, electrical activation, and chemical activation alone (p < 0.05); and (4) approximately 84% of parthenogenetic blastocysts yielded by the optimized protocol were diploid, which was significantly higher than that of electrical activation blastocysts (40%). Using the optimized electrical and combined activation protocol, high blastocyst rates were generated by intracytoplasmic sperm injection (ICSI) (34.6 ± 1.1%), cytoplasmic microinjection (CI) (52.3 ± 2.2%), and handmade cloning (HMC) (31.2 ± 1.0%), respectively. This study concludes that the optimal activation protocol of in vitro matured porcine oocytes was combined activation with parameter as three DC pulses of 1.00 kV cm(-1) for 80 μs plus CB and CHX treatment.

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Section: Introductionmentioning

confidence: 99%

Comparison of chemical, electrical, and combined activation methods for in vitro matured porcine oocytes

Liu

Cui

et al. 2014

In Vitro Cell.Dev.Biol.-Animal

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“…Since MPF activity can suddenly decrease to basal levels after several hours of oocyte activation, the reprogramming of the donor nucleus can only occur in non-activated oocytes. In order to prolong exposure of the donor nucleus to non-activated oocyte, delay activation of reconstructed embryos was applied in previous studies and showed a positive effect on the development of intraspecies cloned embryos (Cibelli et al 1998;Wakayama et al 1998;Wells et al 1998Wells et al , 1999Miyoshi et al 2000;De Sousa et al 2002;Yin et al 2003;Lee et al 2005;Shen et al 2008). Lower efficiency in iSCNT makes it valuable to determine whether longtime exposure of donor nucleus to xenogenic ooplasm benefits the donor nucleus to be reprogrammed and can increase the developmental ability of iSCNT-derived embryos.…”

Section: Discussionmentioning

confidence: 97%

“…Metaphase-II (MII) oocytes have "reprogramming factors" in their cytoplasm, which benefit the reprogramming of donor cells. Some researchers prolonged exposure of incoming nuclei to a non-activated ooplasm (Cibelli et al 1998;Wakayama et al 1998;Wells et al 1998Wells et al , 1999Miyoshi et al 2000;De Sousa et al 2002;Yin et al 2003;Lee et al 2005;Shen et al 2008) or even pretreated donor cells with extract of mitotic somatic cells (Sullivan et al 2004;Rodriguez-Osorio et al 2009;McLean et al 2010) so that the donor nucleus could be well reprogrammed. All of these attempts were performed in intraspecies SCNT and showed a positive effect on the development of cloned embryos.…”

Section: Introductionmentioning

confidence: 98%

Two-staged nuclear transfer can enhance the developmental ability of goat–sheep interspecies nuclear transfer embryos in vitro

Cai

et al. 2010

In Vitro Cell.Dev.Biol.-Animal

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The technique of interspecies somatic cell nuclear transfer, in which interspecies cloned embryos can be reconstructed by using domestic animal oocytes as nuclear recipients and endangered animal or human somatic cells as nuclear donors, can afford more opportunities in endangered animal rescue and human tissue transplantation, but the application of this technique is limited by extremely low efficiency which may be attributed to donor nucleus not fully reprogrammed by xenogenic cytoplasm. In this study, goat fetal fibroblasts (GFFs) were used as nuclear donors, in vitro-matured sheep oocytes were used as nuclear recipients, and a two-stage nuclear transfer procedure was performed to improve the developmental ability of goat-sheep interspecies clone embryos. In the first stage nuclear transfer (FSNT), GFFs were injected into the ooplasm of enucleated sheep metaphase-II oocytes, then non-activated reconstructed embryos were cultured in vitro, so that the donor nucleus could be exposed to the ooplasm for a period of time. Subsequently, in the second stage nuclear transfer, FSNT-derived non-activated reconstructed embryo was centrifuged, and the donor nucleus was then transferred into another freshly enucleated sheep oocyte. Compared with the one-stage nuclear transfer, two-stage nuclear transfer could significantly enhance the blastocyst rate of goat-sheep interspecies clone embryos, and this result indicated that longtime exposure to xenogenic ooplasm benefits the donor nucleus to be reprogrammed. The two-stage nuclear transfer procedure has two advantages, one is that the donor nucleus can be exposed to the ooplasm for a long time, the other is that the problem of oocyte aging can be solved.

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“…Optimalisasi maturasi in vitro oosit antara lain adalah pengklasifikasian oosit, penambahan zat aditif berupa faktor pertumbuhan dan hormon (Shen et al, 2008), maupun berbagai macam serum. Klasifikasi oosit yang didasarkan pada kelengkapan struktur oosit sangat mempengaruhi maturasi in vitro dan perkembangan oosit sampai ke tahap blastosist (Alm et al, 2005;Ksiazkiewicz et al, 2007).…”

Section: Pendahuluanunclassified

Pengaruh Suplementasi Fetal Calf Serum Terhadap Kemampuan Maturasi in Vitro Oosit Sapi

Daoed¹,

Ngadiyono²,

Widayati³

2014

BuletinPeternak

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INTISARIPenelitian ini memanfaatkan hasil samping rumah potong hewan (RPH) sebagai sumber oosit untuk in vitro fertilization (IVF). Untuk meningkatkan keberhasilan IVF dilakukan suplementasi fetal calf serum (FCS) pada medium maturasi in vitro. Ovarium sapi dari RPH dibawa ke laboratorium dalam medium NaCl 0,9% pada suhu 31-34ºC. Selanjutnya oosit diaspirasi menggunakan syringe 3 ml dan jarum 23 G yang berisi Dulbeco's-Phosphate Buffer Saline (DPBS), kemudian dimaturasikan pada inkubator CO 2 modifikasi dalam medium Tissue Culture (CO 2 5%, kelembaban 99% dan suhu 37-39ºC). Oosit dibagi menjadi 2 kelompok yaitu kelompok kontrol (TCM-199) dan kelompok perlakuan (TCM-199 + 10% FCS). Angka maturasi dianalisa dengan Chi-Square, sedangkan kualitas oosit dianalisis secara deskriptif. Maturasi oosit sapi pada kelompok perlakuan berbeda nyata (P≤0,05) dibandingkan kelompok kontrol (55,22% vs 40,09%). Penggunaan 10% FCS pada medium maturasi dapat menghasilkan kualitas oosit matur yang lebih baik dibandingkan kelompok kontrol. Kesimpulan penelitian ini adalah penggunaan 10% FCS suplementasi dapat meningkatkan kemampuan maturasi oosit sapi in vitro. (Kata kunci: Fetal calf serum, Kultur oosit, Maturasi in vitro, Oosit sapi) ABSTRACT This study was conducted to investigate the utilization of ovaries from the slaughterhouse as oocytes source for in vitro fertilization (IVF). Fetal calf serum (FCS) was used as supplement of in-vitro maturation medium in order toincrease the successfullness of IVF. Ovaries were collected from local slaughterhouse and transported to the laboratory using 0.9% NaCl medium at 31-34ºC. The oocytes were aspirated by using a 3 ml syringe and 23 G needle containing Dulbeco's-Phosphate Buffer Saline (DPBS), then were maturated in modified CO 2 incubator (99% humidity and a temperature of 37-39ºC) in Tissue Culture . Oocytes were divided into 2 groups: control group

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The effect of activation treatments on the development of reconstructed bovine oocytes

Cited by 24 publications

References 42 publications

Comparison of chemical, electrical, and combined activation methods for in vitro matured porcine oocytes

Comparison of chemical, electrical, and combined activation methods for in vitro matured porcine oocytes

Two-staged nuclear transfer can enhance the developmental ability of goat–sheep interspecies nuclear transfer embryos in vitro

Pengaruh Suplementasi Fetal Calf Serum Terhadap Kemampuan Maturasi in Vitro Oosit Sapi

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