NMR spectroscopy of a single mammalian early stage embryo

Sivelli, G.; Conley, Gaurasundar M.; Herrera, Carolina; Marable, K.; Rodriguez, Kyle J.; Bollwein, Heinrich; Sudano, Mateus José; Brugger, Jürgen; Simpson, André J.; Boero, Giovanni; Grisi, Marco

doi:10.1016/j.jmr.2021.107142

Cited by 12 publications

(12 citation statements)

References 60 publications

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“…However, advancements in assessing their function are limited. Thirteen studies evaluated embryos 89–101 . Oxygen consumption was investigated by five groups (Figure 5a,b), 89–93 while only one study assessed embryo mechanics 94 .…”

Section: Resultsmentioning

confidence: 99%

“…Cell apoptosis was investigated in one study that assessed the viability 95 . Two studies evaluated embryo structural alterations 100,101 …”

Section: Resultsmentioning

confidence: 99%

“…Thirteen studies evaluated embryos. [89][90][91][92][93][94][95][96][97][98][99][100][101] Oxygen consumption was investigated by five groups (Figure 5a,b), [89][90][91][92][93] while only one study assessed embryo mechanics. 94 Reliable detection of soluble factors at the single-embryo level presents great potential but is challenging.…”

Section: Embryo Evaluationmentioning

confidence: 99%

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Microfluidic chip as a promising evaluation method in assisted reproduction: A systematic review

Wu,

Yan

et al. 2023

Bioengineering & Transla Med

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The aim of assisted reproductive technology (ART) is to select the high‐quality sperm, oocytes, and embryos, and finally achieve a successful pregnancy. However, functional evaluation is hindered by intra‐ and inter‐operator variability. Microfluidic chips emerge as the one of the most powerful tools to analyze biological samples for reduced size, precise control, and flexible extension. Herein, a systematic search was conducted in PubMed, Scopus, Web of Science, ScienceDirect, and IEEE Xplore databases until March 2023. We displayed and prospected all detection strategies based on microfluidics in the ART field. After full‐text screening, 71 studies were identified as eligible for inclusion. The percentages of human and mouse studies equaled with 31.5%. The prominent country in terms of publication number was the USA (n = 13). Polydimethylsiloxane (n = 49) and soft lithography (n = 28) were the most commonly used material and fabrication method, respectively. All articles were classified into three types: sperm (n = 38), oocytes (n = 20), and embryos (n = 13). The assessment contents included motility, counting, mechanics, permeability, impedance, secretion, oxygen consumption, and metabolism. Collectively, the microfluidic chip technology facilitates more efficient, accurate, and objective evaluation in ART. It can even be combined with artificial intelligence to assist the daily activities of embryologists. More well‐designed clinical studies and affordable integrated microfluidic chips are needed to validate the safety, efficacy, and reproducibility.Trial registration: The protocol was registered in the Open Science Frame REGISTRIES (identification: osf.io/6rv4a).

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Section: Resultsmentioning

confidence: 99%

“…Cell apoptosis was investigated in one study that assessed the viability 95 . Two studies evaluated embryo structural alterations 100,101 …”

Section: Resultsmentioning

confidence: 99%

Section: Embryo Evaluationmentioning

confidence: 99%

See 1 more Smart Citation

Microfluidic chip as a promising evaluation method in assisted reproduction: A systematic review

Wu,

Yan

et al. 2023

Bioengineering & Transla Med

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“…Alegre et al proposed Raman spectroscopy, estimating the oxidant content of the culture medium, but its external variability and lower specificity compared to NMR or MS spectroscopy raise concerns [ 4 ]. Hence, NMR emerges as a potential compromise, as demonstrated in recent studies of early-stage mammalian embryos [ 51 ]. Despite evidence supporting metabolic screening’s benefits for nearly a decade, routine implementation in ART clinics is lacking [ 52 , 53 ].…”

Section: Discussionmentioning

confidence: 99%

Metabolomics Integration in Assisted Reproductive Technologies for Enhanced Embryo Selection beyond Morphokinetic Analysis

Pinto,

Guerra-Carvalho,

Crisóstomo

et al. 2023

IJMS

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Embryo quality evaluation during in vitro development is a crucial factor for the success of assisted reproductive technologies (ARTs). However, the subjectivity inherent in the morphological evaluation by embryologists can introduce inconsistencies that impact the optimal embryo choice for transfer. To provide a more comprehensive evaluation of embryo quality, we undertook the integration of embryo metabolomics alongside standardized morphokinetic classification. The culture medium of 55 embryos (derived from 21 couples undergoing ICSI) was collected at two timepoints (days 3 and 5). Samples were split into Good (n = 29), Lagging (n = 19), and Bad (n = 10) according to embryo morphokinetic evaluation. Embryo metabolic performance was assessed by monitoring the variation in specific metabolites (pyruvate, lactate, alanine, glutamine, acetate, formate) using 1H-NMR. Adjusted metabolite differentials were observed during the first 3 days of culture and found to be discriminative of embryo quality at the end of day 5. Pyruvate, alanine, glutamine, and acetate were major contributors to this discrimination. Good and Lagging embryos were found to export and accumulate pyruvate and glutamine in the first 3 days of culture, while Bad embryos consumed them. This suggests that Bad embryos have less active metabolic activity than Good and Lagging embryos, and these two metabolites are putative biomarkers for embryo quality. This study provides a more comprehensive evaluation of embryo quality and can lead to improvements in ARTs by enabling the selection of the best embryos. By combining morphological assessment and metabolomics, the selection of high-quality embryos with the potential to result in successful pregnancies may become more accurate and consistent.

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“…Benefiting from the development of nucleic acid amplification-based sequencing technologies, genomic, transcriptomic, and epigenomic sequencing technologies have been widely used for omic analysis at the single-cell level. , However, it is still a great challenge to analyze proteins and metabolites at the single-cell level due to their limited amount and lack of amplification approaches. Although some tools have been developed and successfully used to analyze some proteins (e.g., mass cytometry and fluorescence imaging) or metabolites (e.g., NMR) in single cells, the numbers of proteins or metabolites analyzed are not sufficient to meet the requirement of omic research. MS has been widely applied in proteomic and metabolomic analysis of the bulk of cellular samples.…”

Section: Introductionmentioning

confidence: 99%

One-Shot Single-Cell Proteome and Metabolome Analysis Strategy for the Same Single Cell

Wu,

Xu,

Jiang

et al. 2024

Anal. Chem.

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Characterizing the profiles of proteome and metabolome at the single-cell level is of great significance in single-cell multiomic studies. Herein, we proposed a novel strategy called one-shot single-cell proteome and metabolome analysis (scPMA) to acquire the proteome and metabolome information in a single-cell individual in one injection of LC-MS/MS analysis. Based on the scPMA strategy, a total workflow was developed to achieve the single-cell capture, nanoliter-scale sample pretreatment, one-shot LC injection and separation of the enzyme-digested peptides and metabolites, and dual-zone MS/MS detection for proteome and metabolome profiling. Benefiting from the scPMA strategy, we realized dual-omic analysis of single tumor cells, including A549, HeLa, and HepG2 cells with 816, 578, and 293 protein groups and 72, 91, and 148 metabolites quantified on average. A single-cell perspective experiment for investigating the doxorubicin-induced antitumor effects in both the proteome and metabolome aspects was also performed.

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NMR spectroscopy of a single mammalian early stage embryo

Cited by 12 publications

References 60 publications

Microfluidic chip as a promising evaluation method in assisted reproduction: A systematic review

Microfluidic chip as a promising evaluation method in assisted reproduction: A systematic review

Metabolomics Integration in Assisted Reproductive Technologies for Enhanced Embryo Selection beyond Morphokinetic Analysis

One-Shot Single-Cell Proteome and Metabolome Analysis Strategy for the Same Single Cell

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