Blastocyst Evaluation by Means of Differential Staining: a Practical Approach

Soom, Ann Van; Vanroose, G.; Kruif, Aart de

doi:10.1046/j.1439-0531.2001.00265.x

Cited by 40 publications

(31 citation statements)

References 73 publications

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“…SCNT embryos activated by PLCZ cRNA injection or common chemical activation stimuli (ionomycin/DMAP and ionomycin/CHX) produced blastocysts at rates of 25 to 36%, which are comparable with those reported in the literature for IVF and SCNT embryos (Cibelli et al 1998, Zakhartchenko et al 1999, Heyman et al 2002, Kane 2003. Total cell number and number of cells allocated to the TE and ICM have been regarded as valuable indicators of cattle embryo quality (van Soom et al 1997(van Soom et al , 2001. A higher total cell number in the embryo correlated with IVF (van Soom et al 1997) and SCNT embryos' developmental potential (Renard et al 2007).…”

Section: Discussionsupporting

confidence: 78%

Activation of bovine somatic cell nuclear transfer embryos by PLCZ cRNA injection

Ross¹,

Rodríguez²,

Iager³

et al. 2009

Reproduction

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The production of cloned animals by the transfer of a differentiated somatic cell into an enucleated oocyte circumvents fertilization. During fertilization, the sperm delivers a sperm-specific phospholipase C (PLCZ) that is responsible for triggering Ca 2C oscillations and oocyte activation. During bovine somatic cell nuclear transfer (SCNT), oocyte activation is artificially achieved by combined chemical treatments that induce a monotonic rise in intracellular Ca 2C and inhibit either phosphorylation or protein synthesis. In this study, we tested the hypothesis that activation of bovine nuclear transfer embryos by PLCZ improves nuclear reprogramming. Injection of PLCZ cRNA into bovine SCNTunits induced Ca 2C oscillations similar to those observed after fertilization and supported high rates of blastocyst development similar to that seen in embryos produced by IVF. Furthermore, gene expression analysis at the eight-cell and blastocyst stages revealed a similar expression pattern for a number of genes in both groups of embryos. Lastly, levels of trimethylated lysine 27 at histone H3 in blastocysts were higher in bovine nuclear transfer embryos activated using cycloheximide and 6-dimethylaminopurine (DMAP) than in those activated using PLCZ or derived from IVF. These results demonstrate that exogenous PLCZ can be used to activate bovine SCNT-derived embryos and support the hypothesis that a fertilization-like activation response can enhance some aspects of nuclear reprogramming.

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Section: Discussionsupporting

confidence: 78%

Activation of bovine somatic cell nuclear transfer embryos by PLCZ cRNA injection

Ross¹,

Rodríguez²,

Iager³

et al. 2009

Reproduction

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“…After incubation, the zona pellucida was removed from each blastocyst with 0.5% pronase (Van Soom et al 2001). Blastocysts were transferred to bisbenzimide dissolved in ethanol (50 mg/ml) for 1 h, washed in 100% ethanol and mounted singly in 1 ml drops of glycerol on siliconised slides.…”

Section: Blastocyst Cell Countsmentioning

confidence: 99%

The effect of paracrine/autocrine interactions on the in vitro culture of bovine preimplantation embryos

Gopichandran

Leese²

2006

Reproduction

114

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Bovine preimplantation embryos develop more successfully when cultured in groups, proibably because of the increased production of, and exposure to, embryotrophic autocrine and paracrine factors. Using a novel embryo culture technique, this study had two aims: 1. to determine the distance over which potential paracrine interactions affect bovine embryo development in terms of blastocyst and hatching rates, cell counts and carbohydrate metabolism; 2. to investigate the effect of platelet-activating factor (PAF) supplementation on bovine embryo development and metabolism. Groups of 16 presumptive zygotes were attached to the bottom of a culture dish by the cell adhesive Cell-Tak in a 4 3 4 equidistant array. The distance between individual embryos in each group was 0-689 mm. Optimal blastocyst formation rate occurred when embryos were cultured 165 mm apart compared with control non-attached zygotes (Kruskal-Wallis followed by Mann-Whitney U test post-hoc; P < 0.05). Increasing the distance between embryos resulted in a further decline in blastocyst rate, which reached zero at 540 mm apart. Blastocyst cell number, pyruvate/glucose uptake and lactate production decreased as the interembryo distance increased from 240 to 465 mm (P < 0.05). Supplementation with PAF during conventional group culture enhanced blastocyst cell number, hatching rates and the oxidative metabolism of pyruvate and glucose. The data indicate that the distance between individual bovine embryos in culture influences preimplantation development, in particular blastocyst formation, cell number and metabolism. It is suggested that diffusible paracrine/autocrine factors, such as PAF, are in part responsible for the regulation of early embryo development.

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“…Embryo evaluation can use invasive and noninvasive techniques [174]. The last group, in assessing embryos without damage, has been widely used based on morphological changes that occur as the result of the sequential cleavage and this classification can be important in preimplantation embryos and especially in embryos that are being transferred [175]. Ushijima et al [176] proposed an 11-stage classification for bovine embryos based on the number of cleavages.…”

Section: Evaluation Of Cryopreserved Embryosmentioning

confidence: 99%

“…Invasive methods of embryo evaluation are used mainly for research proposes, validating the above-described techniques with greater accuracy [175]. Embryo postthawing viability and blastocoele reexpansion in different time periods of embryo development have been widely used to determine its feasibility [13,14,22,181].…”

Section: Evaluation Of Cryopreserved Embryosmentioning

confidence: 99%

Cryopreservation of Sheep Produced Embryos – Current and Future Perspectives

Romão¹,

Marques²,

Bettencourt³

et al. 2016

Insights From Animal Reproduction

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Due to economical and scientific limitations, sheep embryo reproductive technologies are less commercially applied than in other animal species. However, it is very clear that, in the near future, those techniques are expected to have a central role in animal production as a consequence of genetic and reproductive demands. One drawback is that results obtained after sheep embryo cryopreservation are unattractive for commercial purposes. It is expected that a successful cryopreservation of sheep embryos can push forward all other reproductive biotechnologies in this species, such as multiple ovulation and embryo transfer (MOET), artificial insemination, or in vitro production of embryos. This paper tries to discuss the current and future perspectives of cryopreservation of in vivo-and in vitro-produced sheep embryos concerning advantages and limitations for its practical use and possible solutions for improving methods to allow a higher survival rate of cryopreserved embryos.

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Blastocyst Evaluation by Means of Differential Staining: a Practical Approach

Cited by 40 publications

References 73 publications

Activation of bovine somatic cell nuclear transfer embryos by PLCZ cRNA injection

Activation of bovine somatic cell nuclear transfer embryos by PLCZ cRNA injection

The effect of paracrine/autocrine interactions on the in vitro culture of bovine preimplantation embryos

Cryopreservation of Sheep Produced Embryos – Current and Future Perspectives

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