Parthenogenetic activation and subsequent development of rat oocytes in vitro

Jiang, Junfeng; Mizuno, Satoshi; Mizutani, Eiji; Sasada, Hiroshi; Sato, Eimei

doi:10.1002/mrd.1138

Cited by 24 publications

(26 citation statements)

References 53 publications

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“…In goats, a combination treatment of MII oocytes by ethanol and 6-DMAP results in 58% cleavage [22] whereas oocytes reconstructed with somatic cells induce a 56% cleavage [4]. In rats, the percentages of activated oocytes with PN that developed to the 2-cell, 8-cell and blastocyst stages when electrical stimulation was followed by 6-DMAP treatment were significantly higher than the other combined treatments [23]. The rates of 1PN and 1PB formation in buffalo oocytes treated with electric, ethanol or calcium ionophore followed by 6-DMAP were very high (80-100%) whereas the oocytes treated with these agents alone were very low activated (0-20%, unpublished data).…”

Section: Discussionmentioning

confidence: 90%

Development of swamp buffalo (Bubalus bubalis) embryos after parthenogenetic activation and nuclear transfer using serum fed or starved fetal fibroblasts

et al. 2004

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-The knowledge of oocyte activation and somatic cell nuclear transfer in the swamp buffalo (Bubalus bubalis) is extremely rare. The objectives of this study were the following: (1) to investigate the various activation treatments on the parthenogenetic development of buffalo oocytes, (2) to examine the events of nuclear remodeling and in the in vitro development of cloned buffalo embryos reconstructed with serum fed or starved fetal fibroblats, and (3) to investigate the in vivo development of cloned embryos derived from serum fed or starved cells after transfer into the recipients. The rates of cleavage and blastocyst development were found to be significantly higher (P < 0.05) when the oocytes were activated by the combination treatment of calcium ionophore (A23187) or ethanol followed by 6-DMAP than those activated by electrical pulses and 6-DMAP or other single treatments. Flow cytometric analysis revealed that the percentage in the G0/G1 phase in serum starved cells was significantly (P < 0.05) higher than that in serum fed cells (88.8 ± 6.2 vs. 68.2 ± 2.6). At 1 h post fusion (hpf), most of the transferred nuclei (71%) from serum fed cells did not change in size, and the nuclear envelope remained intact, whereas 29% underwent NEBD and PCC. When serum starved cells were used, 83% of the transferred nuclei underwent NEBD and PCC whereas 17% remained intact. The nuclear swelling and pronucleus (PN) formation were observed at 2-4 and 12 h post activation (hpa), respectively. The remodeled nuclei underwent mitotic division and developed to the 2-cell stage within 18-24 hpa. Fifty-five percent of oocytes reconstructed with serum fed cells were 2PN and 45% were 1PN, whereas 79% of the embryos reconstructed from starved cells were 1PN and 21% were 2PN. The percentage of blastocyst development of the embryos derived from starved cells was higher than that from the serum fed cells (35% vs. 21%, P < 0.05). Pregnancy was detected after the transfer of cloned blastocysts into the recipients but no recipients supported the development to term. The results of this work can be used to establish effective activation protocols for buffalo oocytes which can be used during nuclear transfer experiments.buffalo / oocyte activation / nuclear transfer / nuclear remodeling / somatic cell * Corresponding author: scykt@mahidol.ac.th 66 J. Saikhun et al.

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Section: Discussionmentioning

confidence: 90%

Development of swamp buffalo (Bubalus bubalis) embryos after parthenogenetic activation and nuclear transfer using serum fed or starved fetal fibroblasts

et al. 2004

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“…In a previous study, we reported that the combination of electrical stimulation with 6-DMAP markedly improved parthenogenetic activation and subsequent development of rat oocytes to the blastocyst stage in vitro [10]. However, the percentage of blastocysts developed from activated rat oocytes wa s still low and the quality of blastocysts obtained was poor.…”

Section: Discussionmentioning

confidence: 91%

“…Animals were given bullet-type commercial rat feed [High Pure Ace (P), Itochu Co., Ishinomaki, Japan) and tap water ad libitum, as described previously [10]. The studies were approved by the Ethics Committee for Care and Use of Laboratory Animals for Biomedical Research of the Graduate School of Agricultural Science, Tohoku University.…”

Section: Preparation Of Animalsmentioning

confidence: 99%

“…Oocytes were activated as described previously [10]. Briefly, after washing, oocytes were placed between two wire electrodes (1 mm apart) on a fusion chamber slide with 30 ml of activation solution containing mannitol, 0.1 mM CaCl 2 , 0.1 mM MgSO4 (all from Wako) and fatty acid-free BSA (Sigma).…”

Section: Oocyte Activation By Electrical Stimulation Combined With 6-mentioning

confidence: 99%

“…Our recent results showed that 50% of rat oocytes developed to blastocysts when activation was induced by e l e c t r i c a l s ti m u l a t i o n c o m b i n e d w i t h 6 -dim ethylaminopurine (6-DMAP) treatment. However, the quality of these blastocysts was poor in terms of morphology and the cell number was low (26.9 ± 1.9), ranging from 22 to 36 [10]. A more recent study showed that Wistar rat oocytes could be activated by electrical stimulation, ethanol or strontium treatment.…”

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confidence: 99%

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Determination of Optimal Conditions for Parthenogenetic Activation and Subsequent Development of Rat Oocytes In Vitro

Mizutani

Jiang

Mizuno

et al. 2004

Journal of Reproduction and Development

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Abstract. The present study was undertaken to determine optimal conditions for parthenogenetic activation and subsequent development of rat oocytes. Oocytes from immature Wistar-Imamichi (WI) and Sprague Dawley (SD) rats were activated by electrical stimulation in combination with 6-dimethylaminopurine (6-DMAP) to assess whether different rat strains display different responses to activation treatment. Since the cleavage rates of activated oocytes were significantly higher in WI than SD strain rats, WI rats were used for the subsequent experiments to determine the effects of post-hCG time, culture duration, different activation protocols (electrical stimulation with 6-DMAP or ionomycin with 6-DMAP) and osmolarity of the activation medium on the activation and subsequent development of WI rat oocytes. For oocytes activated by electrical stimulation combined with 6-DMAP, the percentages of oocytes that were activated and that developed to blastocysts were higher when oocytes were collected at 18-20 h than at any other time points after hCG injection (16, 22-24 h). Culturing for 2-6 h before activation treatment markedly decreased the percentage of activated oocytes that developed to beyond the four-cell stage. There were no differences in the percentages of oocytes with pronuclear formation and subsequent development to the two-cell and blastocyst stages between oocytes that were activated by electrical stimulation or ionomycin, both followed by 6-DMAP treatment. Activation of oocytes by ionomycin and 6-DMAP, both in low osmolarity media (246 mOsM), markedly increased the cleavage rates and percentages of high quality blastocysts (71%). The optimal conditions determined in the present study with simplified activation protocols and high efficiency of activation and subsequent development of WI rat oocytes will be helpful for further research involving nuclear transfer in the rat.

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Two‐phase chemically defined culture system for preimplantation rat embryos

Yi-jie

Galat

Garton

et al. 2003

Genesis

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A limiting factor in the development of new technologies and transport of rats worldwide has been the inability to robustly culture preimplantation embryos. Previously, culture in vitro to the blastocyst stage from one-cell embryos was successful only if the one-cell embryos were isolated near the time of the first cleavage and from only a few strains. Here we report the use of commonly available, chemically defined culture media to overcome these limitations. In vitro culture of young one-cell embryos using common embryo media (KSOM, BMOC, or HTF) for 18-22 h followed by culture in mR1ECM medium allows the successful in vitro development of blastocysts from one-cell embryos after 5 days from both outbred (SD) and inbred strains of rat (WF, LEW, F344, and PVG). This system allows the parthenogenetic development of chemically activated, unfertilized oocytes to the blastocyst stage. Embryos cultured in this system develop to term and are live-born following transfer to surrogate mothers.

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Parthenogenetic activation and subsequent development of rat oocytes in vitro

Cited by 24 publications

References 53 publications

Development of swamp buffalo (Bubalus bubalis) embryos after parthenogenetic activation and nuclear transfer using serum fed or starved fetal fibroblasts

Development of swamp buffalo (Bubalus bubalis) embryos after parthenogenetic activation and nuclear transfer using serum fed or starved fetal fibroblasts

Determination of Optimal Conditions for Parthenogenetic Activation and Subsequent Development of Rat Oocytes In Vitro

Two‐phase chemically defined culture system for preimplantation rat embryos

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