Using Time Lapse Monitoring for Determination of Morphological Defect Frequency in Feline Embryos after in Vitro Fertilization (IVF)

Kij, Barbara; Kochan, Joanna; Nowak, Agnieszka; Niżański, Wojciech; Prochowska, Sylwia; Fryc, Karolina; Bugno‐Poniewierska, Monika

doi:10.3390/ani10010003

Cited by 8 publications

(6 citation statements)

References 26 publications

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“…In our previous study we determined the incidence of morphological defects in feline embryos [ 22 ]. We divided defects into two groups: single or multiple aberration.…”

Section: Discussionmentioning

confidence: 99%

“…According to our previous research [ 22 ], embryos with morphological disorders have the potential to reach the blastocyst stage as well as normal embryos but are less likely to hatch. The hatching rate was the highest in the normally cleaving embryos (15.6%) and decreased significantly within the groups that exhibited a single aberration (6.25%) and multiple aberrations (3.33%).…”

Section: Discussionmentioning

confidence: 99%

“…In cats, the association between the onset of the first cleavage division and blastocyst formation has only been studied using traditional microscopic observation of embryos at 18, 24 and 30 h post-insemination [ 12 ] or 27 and 42 h post-insemination [ 21 ]. In our previous pilot studies [ 22 ], we used time lapse to monitor feline embryos for the first time and determined the frequency of morphological defects of embryos and their competence to reach the blastocyst stage, as well as their ability to hatch. Now, as a continuation of that research, we have made an attempt to precisely define morphokinetic parameters in feline embryos using a time lapse system (Primo Vision ® ).…”

Section: Introductionmentioning

confidence: 99%

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Analysis of Morphokinetic Parameters of Feline Embryos Using a Time-Lapse System

Kochan

Nowak

Kij

et al. 2021

Animals

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The aim of this study was to analyze the morphokinetic parameters of feline embryos using a time lapse system. Oocytes matured in vitro were fertilized (IVF) and in vitro cultured in a time lapse-system (Primo Vision®, Gothenburg, Sweden). The first cell division of embryos occurred between 17 h post insemination (hpi) and 38 hpi, with the highest proportion of embryos (46%) cleaving between 21 and 24 hpi. The timing of the first cleavage significantly affected further embryo development, with the highest development occurring in embryos that cleaved at 21–22 hpi. Embryos that cleaved very early (17–18 hpi) developed poorly to the blastocyst stage (2%) and none of the embryos that cleaved later than 27 hpi were able to reach the blastocyst stage. Morphological defects were observed in 48% of the embryos. There were no statistically significant differences between the timing intervals of the first cleavage division and the frequency of morphological defects in embryos. Multiple (MUL) morphological defects were detected in more than half (56%) of the abnormal embryos. The most frequent single morphological defects were cytoplasmic fragmentation (FR) (8%) and blastomere asymmetry (AS) (6%). Direct cleavage (DC) from 1–3 or 3–5 blastomeres, reverse cleavage (RC) and vacuoles were rarely observed (2–3%). The timing of blastocyst cavity formation is a very good indicator of embryo quality. In our study, blastocyst cavity formation occurred between 127–167 hpi, with the highest frequency of hatching observed in blastocysts that cavitated between 142–150 hpi. Blastocysts in which cavitation began after 161 h did not hatch. In conclusion, the timing of the first and second cleavage divisions, the timing of blastocyst cavity formation and morphological anomalies can all be used as early and non-invasive indicators of cat embryo development in vitro.

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“…In our previous study we determined the incidence of morphological defects in feline embryos [ 22 ]. We divided defects into two groups: single or multiple aberration.…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Analysis of Morphokinetic Parameters of Feline Embryos Using a Time-Lapse System

Kochan

Nowak

Kij

et al. 2021

Animals

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“…After removing the connective tissues, fat mass, and fallopian tubes, the ovaries were soaked in 75% alcohol for 30 s and washed 3 times with a 0.9% sodium chloride solution preheated to 38.5 °C. The ovaries were placed into a 60-mm petri dish containing TCM-199 medium [ 24 ] with 6 mg/mL BSA (washing medium) and fixed with tweezers. Subsequently, the follicles on the surface of the ovary were lacerated with a scalpel.…”

Section: Methodsmentioning

confidence: 99%

TLR7/8 signalling affects X-sperm motility via the GSK3 α/β-hexokinase pathway for the efficient production of sexed dairy goat embryos

Ren

Hao

Ren

et al. 2021

J Animal Sci Biotechnol

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Background Goat milk is very similar to human milk in terms of its abundant nutrients and ease of digestion. To derive greater economic benefit, farmers require more female offspring (does); however, the buck-to-doe offspring sex ratio is approximately 50%. At present, artificial insemination after the separation of X/Y sperm using flow cytometry is the primary means of controlling the sex of livestock offspring. However, flow cytometry has not been successfully utilised for the separation of X/Y sperm aimed at sexing control in dairy goats. Results In this study, a novel, simple goat sperm sexing technology that activates the toll-like receptor 7/8 (TLR7/8), thereby inhibiting X-sperm motility, was investigated. Our results showed that the TLR7/8 coding goat X-chromosome was expressed in approximately 50% of round spermatids in the testis and sperm, as measured from cross-sections of the epididymis and ejaculate, respectively. Importantly, TLR7/8 was located at the tail of the X-sperm. Upon TLR7/8 activation, phosphorylated forms of glycogen synthase kinase α/β (GSK3 α/β) and nuclear factor kappa-B (NF-κB) were detected in the X-sperm, causing reduced mitochondrial activity, ATP levels, and sperm motility. High-motility Y-sperm segregated to the upper layer and the low-motility X-sperm, to the lower layer. Following in vitro fertilisation using the TLR7/8-activated sperm from the lower layer, 80.52 ± 6.75% of the embryos were XX females. The TLR7/8-activated sperm were subsequently used for in vivo embryo production via the superovulatory response; nine embryos were collected from the uterus of two does that conceived. Eight of these were XX embryos, and one was an XY embryo. Conclusions Our study reveals a novel TLR7/8 signalling mechanism that affects X-sperm motility via the GSK3 α/β-hexokinase pathway; this technique could be used to facilitate the efficient production of sexed dairy goat embryos.

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“…However, further research resulted in the introduction of sequential culture media, which were based on the composition of human tubal uid [19]. Then, with the introduction of time-lapse technology in embryology and some new clinical trials, single-step culture media gained renewed attention and were started to be used routinely in embryo culture media, especially when time-lapse technology was used to monitor embryos [20][21][22][23]. Single-step media can also be useful in the zona-free somatic cell nuclear transfer (SCNT) method, in which the embryos should be cultured in an undisturbed environment to support the development of SCNT embryos to the blastocyst stage.…”

Section: Introductionmentioning

confidence: 99%

The Impact of Two Embryo Culture Media, SOF and Commercial BO, on Pre- and Post-Implantation Development of Cloned Sannen Goat Embryos

Hajian

Jafarpour

Aghamiri

et al. 2020

Preprint

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Background: The ingredients of embryo culture media developed by different companies are disclosed. Thus, it is impossible to determine which ingredients might be responsible for differences in pre-and post-implantation embryo development. To address this gap, we performed an experiment to compare two embryo culture media, namely, SOF and commercial BO, on pre- and post-implantation development of cloned Sannen goat embryos. Cumulus oocyte complexes derived from slaughterhouse ovaries were used for in vitro embryo production . In vitro development of IVF, parthenogenetic and SCNT embryos were assessed in both BO and SOF media. The expression of 16 genes, including AKT , OCT4 , SOX2 , BMPR1 , FGFR4 , CDC25 , CDX2 , GCN5 , PCAF , FOXD3 , SMAD5 , FZD , LIFR1 , CTNNB , ERK1 , and IFNT , belonging to 7 important pathways, i.e. pluripotency, FGF, TGFβ, cell cycle and proliferation, histone transferase, trophectoderm, and WNT, were examined in the goat SCNT and IVF blastocysts from both BO and SOF media. Results: The blastocyst rate in BO medium was significantly higher than that of the SOF medium in SCNT embryos ( P < 0.05). All of the genes examined showed increased expression levels in SCNT embryos compared to IVF embryos. In the IVF group, OCT4 , BMPR1 , and GCN5 showed significantly higher expression in the SOF medium compared to the BO medium. In this group, AKT , FGFR4 , SOX2 showed significantly lower expression in the SOF medium compared to the BO medium. In the SCNT group, FGFR4 , GCN5 , FZD , CTNNB , BMPR1 , and FGFR4 showed significantly higher expression in SOF medium compared to BO medium. In vivo development did not differ significantly between the two groups. Conclusions: Based on these results, we concluded that the limited information available on the allocations of ICM and TE cells in SCNT embryos and embryo-specific gene expression may be the major drawback IVC medium and an impediment to successful animal cloning.

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Using Time Lapse Monitoring for Determination of Morphological Defect Frequency in Feline Embryos after in Vitro Fertilization (IVF)

Cited by 8 publications

References 26 publications

Analysis of Morphokinetic Parameters of Feline Embryos Using a Time-Lapse System

Analysis of Morphokinetic Parameters of Feline Embryos Using a Time-Lapse System

TLR7/8 signalling affects X-sperm motility via the GSK3 α/β-hexokinase pathway for the efficient production of sexed dairy goat embryos

The Impact of Two Embryo Culture Media, SOF and Commercial BO, on Pre- and Post-Implantation Development of Cloned Sannen Goat Embryos

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