The use of green ﬂuorescent protein (GFP)
to select bovine embryos

Duszewska, A. M.; Козикова, Л. В.; Szydlik, H.; Cybulska, Magdalena; Korwin-Kossakowski, M.; Wąs, Bogdan; Połoszynowicz, J.; Wicińska, K.; Rosochacki, S. J.

doi:10.22358/jafs/67661/2003

Cited by 3 publications

(4 citation statements)

References 31 publications

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“…Transient expression was detected in 2 out of 357 cleaved embryos and these were removed from in vitro culture. Transient expression can result from episomal expression of unintegrated DNA and is a major barrier in the generation of transgenic animals (Wall, 1997;Duszewska et al, 2004) The effi ciency of production of GFP-positive embryos (3.60%) was lower than in our previous study, 6.57% (Duszewska, 2003) and also in comparison with other studies: 6% (Eyestone, 1999) and 14% (Behboodi et al, 2001).…”

Section: Discussioncontrasting

confidence: 52%

“…The embryos from microinjected and non-injected groups were co-cultured with Vero cells which support embryo development better than other systems of culture used in our laboratory (Duszewska et al, 2000(Duszewska et al, , 2003. Microinjection is a traumatic procedure for zygotes and can cause lysis or reduce early embryo development (Han et al, 2000;Chan et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

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Obtaining calves after transfer of embryos microinjected with the human interferon alpha (IFNα) gene, pbLGIFN-GFPBsd

Duszewska¹,

Lipiński²,

Gawron³

et al. 2008

J. Anim. Feed Sci.

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The objective of this study was to obtain calves with an integrated human interferon alpha (IFNα) gene. The transgene (pbLGIFN-GFPBsd) was introduced by microinjection into one of the zygote's pronuclei. The transgene (pbLGIFN-GFPBsd) contained the human interferon alpha (IFNα) gene, bovine lactoglobulin promoter (bLG), gfp (green fl uorescence protein) and blasticidin resistance gene (bsd). After microinjection of 2107 zygotes, the proportion developing to the blastocyst stage (10.4%) was lower than for non-injected embryos (39.9%; P<0.01). Of the 2107 that were microinjected, 76 were GFP-positive blastocysts (3.6%). Fifty-seven GFP-positive blastocysts and 31 blastocysts from non-injected embryos were transferred singly to recipients. A lower rate of calving was observed after transfer of GFP-positive blastocysts (19.3%) as compared with noninjected embryos (48.4%; P<0.05). All newborn calves were healthy and of normal weight. PCR analyses indicated that none of the calves obtained after transfer of GFP-positive blastocysts carried the human interferon alpha gene.

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Section: Discussioncontrasting

confidence: 52%

Section: Discussionmentioning

confidence: 99%

Obtaining calves after transfer of embryos microinjected with the human interferon alpha (IFNα) gene, pbLGIFN-GFPBsd

Duszewska¹,

Lipiński²,

Gawron³

et al. 2008

J. Anim. Feed Sci.

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“…In farm animals, these long-term consequences lead to abnormal birth weight and postnatal growth abnormality, referred to as Large Offspring Syndrome (LOS) (van Wagtgendonk et al, 2000;Bertolini et al, 2002). Sometimes, but not usually, LOS has been associated with in vitro production of embryos, including nuclear transfer and pronuclear injection (Young and Fairburn 2000;review: van Wagtgendonk et al, 2000;Renard et al, 2002;Duszewska et al, 2003Duszewska et al, , 2004.…”

Section: Discussionmentioning

confidence: 99%

The development potential of bovine embryo co-culture with Vero and Vero/BRL cells

Duszewska¹,

Wojdan²,

Gawron³

et al. 2007

J. Anim. Feed Sci.

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The objective of this study was to evaluate the effect of two bovine embryo co-culture systems on calving rates. The embryos were co-cultured with Vero or Vero/BRL cells (mixed co-culture) until the blastocyst stage. A higher percentage of blastocysts was observed on Vero/BRL cells, 42.85%, compared with 30.41% on Vero cells (P≤0.05). The blastocysts from Vero/BRL and Vero cells were transferred to recipients. A higher rate of calving was obtained after transfer of embryos co-cultured with Vero cells (37.50%), compared with Vero/BRL cells (13.16%) (P≤0.01). A greater loss of pregnancy after transfer of embryos co-cultured with Vero/BRL cells was observed between 35 and 65 days. All calves were born naturally, healthy, and with normal weight. Vero cells better support cattle prenatal development than Vero/BRL cells. Probably Vero/BRL cells lead to epigenetic modifications that are responsible for early foetal resorption.

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“…Visualization of pronuclei [30] Evaluation of embryos between days 4 and 8 of culture [31] Evaluation of cleavage [32] Visualization of lipid droplets [33] Comparison of 8-cell-stage embryos in different culture media and visualization of cytoplasmic lipid droplets [34] Visualization of DNA microinjection into the male pronucleus [35,36] Observation of the development of individual embryos [37] Observation of cytoplasmic lipid droplets [38,39] 3.1.…”

Section: Applications Of Dic In Bovine Embryo Analysis Sourcementioning

confidence: 99%

Pre-Implantation Bovine Embryo Evaluation—From Optics to Omics and Beyond

Rabel

Marchioretto

Bangert

et al. 2023

Animals

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Approximately 80% of the ~1.5 million bovine embryos transferred in 2021 were in vitro produced. However, only ~27% of the transferred IVP embryos will result in live births. The ~73% pregnancy failures are partly due to transferring poor-quality embryos, a result of erroneous stereomicroscopy-based morphological evaluation, the current method of choice for pre-transfer embryo evaluation. Numerous microscopic (e.g., differential interference contrast, electron, fluorescent, time-lapse, and artificial-intelligence-based microscopy) and non-microscopic (e.g., genomics, transcriptomics, epigenomics, proteomics, metabolomics, and nuclear magnetic resonance) methodologies have been tested to find an embryo evaluation technique that is superior to morphologic evaluation. Many of these research tools can accurately determine embryo quality/viability; however, most are invasive, expensive, laborious, technically sophisticated, and/or time-consuming, making them futile in the context of in-field embryo evaluation. However accurate they may be, using complex methods, such as RNA sequencing, SNP chips, mass spectrometry, and multiphoton microscopy, at thousands of embryo production/collection facilities is impractical. Therefore, future research is warranted to innovate field-friendly, simple benchtop tests using findings already available, particularly from omics-based research methodologies. Time-lapse monitoring and artificial-intelligence-based automated image analysis also have the potential for accurate embryo evaluation; however, further research is warranted to innovate economically feasible options for in-field applications.

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The use of green ﬂuorescent protein (GFP) to select bovine embryos

Cited by 3 publications

References 31 publications

Obtaining calves after transfer of embryos microinjected with the human interferon alpha (IFNα) gene, pbLGIFN-GFPBsd

Obtaining calves after transfer of embryos microinjected with the human interferon alpha (IFNα) gene, pbLGIFN-GFPBsd

The development potential of bovine embryo co-culture with Vero and Vero/BRL cells

Pre-Implantation Bovine Embryo Evaluation—From Optics to Omics and Beyond

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