Non-invasive assessment of in-vitro embryo quality to improve transfer success

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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Respiration rates of individual bovine in vitro-produced embryos measured with a novel, non-invasive and highly sensitive microsensor system

Reproduction ◽

10.1530/rep.1.00703 ◽

2005 ◽

Vol 130 (5) ◽

pp. 669-679 ◽

Cited By ~ 66

Author(s):

A S Lopes ◽

L H Larsen ◽

N Ramsing ◽

P Løvendahl ◽

M Räty ◽

...

Keyword(s):

Respiration Rate ◽

Embryo Quality ◽

Developmental Stages ◽

Embryo Morphology ◽

The Novel ◽

Non Invasive ◽

Respiration Rates ◽

Morphological Evaluation ◽

Highly Sensitive

Oxygen consumption is a useful parameter for evaluating embryo quality, since it provides a valuable indication of overall metabolic activity. Over the years, several approaches have been used to measure the respiration rates of individual embryos, but a convincing method has not yet been reported. In this study, we introduce and have validated a novel high resolution microsensor technology to determine the respiration rates of individual embryos at different developmental stages. We have employed this technology to investigate the correlation between respiration rate and embryo morphology, diameter and sex. Following morphological evaluation, individual respiration rates of day 3 (n = 18) and day 7 (n = 60) bovine in vitro-produced embryos were determined. Of the measured embryos, 64 were lysed for sex diagnosis by PCR. Average respiration rates of day 7 embryos (1.30 ± 0.064 nl/h) were 3.4-fold higher than day 3 embryos (0.38 ± 0.011 nl/h). On day 7, the average respiration rate of quality 1 blastocysts was significantly higher than the respiration rates of the lower qualities. For both day 3 and day 7 embryos, respiration rates were directly influenced by embryo diameter but did not differ between sexes. These results have demonstrated that the novel microsensor technology can be used to accurately and rapidly (8 min) measure the respiration rates of individual embryos at different developmental stages. Respiration rates were only in partial agreement with embryo morphology, suggesting a slight discrepancy between these two methods in assessing embryo quality. It is likely that a combined assessment of embryo respiration and morphology would improve embryo classification and subsequent selection.

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The invasive and new non-invasive methods of mammalian oocyte and embryo quality assessment: a review

Veterinární Medicína ◽

10.17221/5913-vetmed ◽

2012 ◽

Vol 57 (No. 4) ◽

pp. 169-176 ◽

Cited By ~ 2

Author(s):

D. Bukowska ◽

B. Kempisty ◽

H. Piotrowska ◽

R. Walczak ◽

P. Sniadek ◽

...

Keyword(s):

Quality Assessment ◽

Embryo Quality ◽

Developmental Potential ◽

Vitro Fertilization ◽

Lab On Chip ◽

New Device ◽

Non Invasive ◽

Mammalian Oocyte ◽

On Chip

 The quality of oocytes-embryos can be determined by several techniques, including morphological, molecular, cellular and biochemical ones. The morphological methods of female gamete or embryo quality assessment often use thе following in vitro manipulation procedures such as: in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro embryo production (IVP). However, these methods are highly subjective and the morphological classification of oocytes or embryos is not always compatible with their ability to grow and develop. Additionally, molecular biology methods are objective and present parametric results, which are more or less comparable to the real oocyte-embryo “health”. Although these techniques enable us to determine markers of oocyte-embryo developmental potential, when applied they lead to destruction of the analysed cells. Therefore, the need still exists to search for new methods that will be highly objective (parametric) and, which is most important, non-invasive. In this review, the morphological and molecular methods of oocyte-embryo quality assessment are presented. Moreover, we described a new system based on microfluidic technology (Lab-on-Chip) which allows the creation of a new device for mammalian oocyte as well as embryo quality evaluation: by using their spectral characterisation following embryo transfer (ET) procedures in the cattle and the pig.    

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Non-invasive biomarkers for embryo quality in sterile patients

10.26226/morressier.5af300af738ab10027aa9499 ◽

2018 ◽

Author(s):

Iris Martínez Rodero ◽

Juan Manuel Moreno Moya ◽

Josep Marí-Alexandre ◽

Irene Cuevas Saiz ◽

Carla Olmedo Illueca

Keyword(s):

Embryo Quality ◽

Non Invasive

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Follicular fluid and assisted reproductive technology programs outcomes (literature review)

GYNECOLOGY ◽

10.26442/20795696.2019.6.190663 ◽

2020 ◽

Vol 21 (6) ◽

pp. 36-40

Author(s):

Anna G. Burduli ◽

Natalia A. Kitsilovskaya ◽

Yuliya V. Sukhova ◽

Irina A. Vedikhina ◽

Tatiana Y. Ivanets ◽

...

Keyword(s):

Clinical Practice ◽

Assisted Reproductive Technology ◽

Follicular Fluid ◽

Economic Cost ◽

Reproductive Technology ◽

Art Programs ◽

Vitro Fertilization ◽

Non Invasive ◽

Established Fact

The review presents data on metabolites in the follicular fluid (FF) from the perspective of reproductive medicine and their use in order to predict outcomes of assisted reproductive technology (ART) programs. It considers various components of this biological medium (hormones, lipids, melatonin, etc.) with an assessment of their predictive value in prognosis of the effectiveness of in vitro fertilization (IVF) programs. The data on experimental directions in this field and the prospects for their use in clinical practice are presented. The article emphasizes that the growing clinical need and the unsolved problem of increasing the effectiveness of ART programs determine the need for further studies of the FF composition. Materials and methods. The review includes data related to this topic from foreign and Russian articles found in PubMed which were published in recent years. Results. Given the established fact of a direct effect of FF composition on growth and maturation of oocytes, and further, on the fertilization process, various FF metabolites are actively investigated as non-invasive markers of quality of oocytes/embryos. The article provides data on the experimental directions in this field and the prospects for their use in clinical practice. However, clinical studies of a relation between various FF metabolites levels and outcomes of IVF programs are contradictory. Conclusion. Owing large economic cost for treatment of infertility with IVF, there is need for expansion and intensification of studies to identify and use reliable predictors in prognosis of ART programs outcomes.

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Picosecond acoustics in vegetal cells: Non-invasive in vitro measurements at a sub-cell scale

Ultrasonics ◽

10.1016/j.ultras.2009.09.019 ◽

2010 ◽

Vol 50 (2) ◽

pp. 202-207 ◽

Cited By ~ 22

Author(s):

B. Audoin ◽

C. Rossignol ◽

N. Chigarev ◽

M. Ducousso ◽

G. Forget ◽

...

Keyword(s):

Non Invasive ◽

In Vitro Measurements

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Non-invasive skin sampling of tryptophan/kynurenine ratio in vitro towards a skin cancer biomarker

Scientific Reports ◽

10.1038/s41598-020-79903-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Skaidre Jankovskaja ◽

Johan Engblom ◽

Melinda Rezeli ◽

György Marko-Varga ◽

Tautgirdas Ruzgas ◽

...

Keyword(s):

Skin Cancer ◽

Relative Increase ◽

Cancer Biomarker ◽

Cancer Diagnostics ◽

Sampling Time ◽

Skin Permeability ◽

Experimental Conditions ◽

Non Invasive

AbstractThe tryptophan to kynurenine ratio (Trp/Kyn) has been proposed as a cancer biomarker. Non-invasive topical sampling of Trp/Kyn can therefore serve as a promising concept for skin cancer diagnostics. By performing in vitro pig skin permeability studies, we conclude that non-invasive topical sampling of Trp and Kyn is feasible. We explore the influence of different experimental conditions, which are relevant for the clinical in vivo setting, such as pH variations, sampling time, and microbial degradation of Trp and Kyn. The permeabilities of Trp and Kyn are overall similar. However, the permeated Trp/Kyn ratio is generally higher than unity due to endogenous Trp, which should be taken into account to obtain a non-biased Trp/Kyn ratio accurately reflecting systemic concentrations. Additionally, prolonged sampling time is associated with bacterial Trp and Kyn degradation and should be considered in a clinical setting. Finally, the experimental results are supported by the four permeation pathways model, predicting that the hydrophilic Trp and Kyn molecules mainly permeate through lipid defects (i.e., the porous pathway). However, the hydrophobic indole ring of Trp is suggested to result in a small but noticeable relative increase of Trp diffusion via pathways across the SC lipid lamellae, while the shunt pathway is proposed to slightly favor permeation of Kyn relative to Trp.

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Transcriptional Profiling of Porcine Blastocysts Produced In Vitro in a Chemically Defined Culture Medium

Animals ◽

10.3390/ani11051414 ◽

2021 ◽

Vol 11 (5) ◽

pp. 1414

Author(s):

Josep M. Cambra ◽

Emilio A. Martinez ◽

Heriberto Rodriguez-Martinez ◽

Maria A. Gil ◽

Cristina Cuello

Keyword(s):

Culture Medium ◽

Embryo Quality ◽

Transcriptional Profiling ◽

Defined Medium ◽

Platelet Factor ◽

Defined Media ◽

Defined Culture ◽

The Impact

The development of chemically defined media is a growing trend in in vitro embryo production (IVP). Recently, traditional undefined culture medium with bovine serum albumin (BSA) has been successfully replaced by a chemically defined medium using substances with embryotrophic properties such as platelet factor 4 (PF4). Although the use of this medium sustains IVP, the impact of defined media on the embryonic transcriptome has not been fully elucidated. This study analyzed the transcriptome of porcine IVP blastocysts, cultured in defined (PF4 group) and undefined media (BSA group) by microarrays. In vivo-derived blastocysts (IVV group) were used as a standard of maximum embryo quality. The results showed no differentially expressed genes (DEG) between the PF4 and BSA groups. However, a total of 2780 and 2577 DEGs were detected when comparing the PF4 or the BSA group with the IVV group, respectively. Most of these genes were common in both in vitro groups (2132) and present in some enriched pathways, such as cell cycle, lysosome and/or metabolic pathways. These results show that IVP conditions strongly affect embryo transcriptome and that the defined culture medium with PF4 is a guaranteed replacement for traditional culture with BSA.

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Design and Analysis of a Continuous and Non-Invasive Multi-Wavelength Optical Sensor for Measurement of Dermal Water Content

Sensors ◽

10.3390/s21062162 ◽

2021 ◽

Vol 21 (6) ◽

pp. 2162

Author(s):

Mohammad Mamouei ◽

Subhasri Chatterjee ◽

Meysam Razban ◽

Meha Qassem ◽

Panayiotis A. Kyriacou

Keyword(s):

Water Content ◽

Optical Sensor ◽

Broad Band ◽

Gravimetric Analysis ◽

Skin Damage ◽

Biophysical Parameter ◽

Non Invasive ◽

Multi Wavelength ◽

Invasive Measurement

Dermal water content is an important biophysical parameter in preserving skin integrity and preventing skin damage. Traditional electrical-based and open-chamber evaporimeters have several well-known limitations. In particular, such devices are costly, sizeable, and only provide arbitrary outputs. They also do not permit continuous and non-invasive monitoring of dermal water content, which can be beneficial for various consumer, clinical, and cosmetic purposes. We report here on the design and development of a digital multi-wavelength optical sensor that performs continuous and non-invasive measurement of dermal water content. In silico investigation on porcine skin was carried out using the Monte Carlo modeling strategy to evaluate the feasibility and characterize the sensor. Subsequently, an in vitro experiment was carried out to evaluate the performance of the sensor and benchmark its accuracy against a high-end, broad band spectrophotometer. Reference measurements were made against gravimetric analysis. The results demonstrate that the developed sensor can deliver accurate, continuous, and non-invasive measurement of skin hydration through measurement of dermal water content. Remarkably, the novel design of the sensor exceeded the performance of the high-end spectrophotometer due to the important denoising effects of temporal averaging. The authors believe, in addition to wellbeing and skin health monitoring, the designed sensor can particularly facilitate disease management in patients presenting diabetes mellitus, hypothyroidism, malnutrition, and atopic dermatitis.

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Non-invasive plasma glycomic and metabolic biomarkers of post-treatment control of HIV

Nature Communications ◽

10.1038/s41467-021-24077-w ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Leila B. Giron ◽

Clovis S. Palmer ◽

Qin Liu ◽

Xiangfan Yin ◽

Emmanouil Papasavvas ◽

...

Keyword(s):

Treatment Interruption ◽

Post Treatment ◽

Machine Learning Algorithms ◽

Virologic Suppression ◽

Hiv Latency ◽

Viral Rebound ◽

Treatment Control ◽

Non Invasive ◽

Metabolic Biomarkers

AbstractNon-invasive biomarkers that predict HIV remission after antiretroviral therapy (ART) interruption are urgently needed. Such biomarkers can improve the safety of analytic treatment interruption (ATI) and provide mechanistic insights into the host pathways involved in post-ART HIV control. Here we report plasma glycomic and metabolic signatures of time-to-viral-rebound and probability-of-viral-remission using samples from two independent cohorts. These samples include a large number of post-treatment controllers, a rare population demonstrating sustained virologic suppression after ART-cessation. These signatures remain significant after adjusting for key demographic and clinical confounders. We also report mechanistic links between some of these biomarkers and HIV latency reactivation and/or myeloid inflammation in vitro. Finally, machine learning algorithms, based on selected sets of these biomarkers, predict time-to-viral-rebound with 74% capacity and probability-of-viral-remission with 97.5% capacity. In summary, we report non-invasive plasma biomarkers, with potential functional significance, that predict both the duration and probability of HIV remission after treatment interruption.

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