scholarly journals Nuclear and mitochondrial DNA in blastocoele fluid and embryo culture medium: evidence and potential clinical use

2016 ◽  
Vol 31 (8) ◽  
pp. 1653-1661 ◽  
Author(s):  
Elizabeth R. Hammond ◽  
Andrew N. Shelling ◽  
Lynsey M. Cree
Keyword(s):  
Mitochondrial Dna ◽  
Embryo Culture ◽  
Culture Medium ◽  
Clinical Use ◽  
Embryo Culture Medium

scholarly journals Mitochondrial DNA in Day 3 embryo culture medium is a novel, non-invasive biomarker of blastocyst potential and implantation outcome

2014 ◽  
Vol 20 (12) ◽  
pp. 1238-1246 ◽  
Author(s):  
S. Stigliani ◽  
L. Persico ◽  
C. Lagazio ◽  
P. Anserini ◽  
P.L. Venturini ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Embryo Culture ◽  
Culture Medium ◽  
Non Invasive ◽  
Embryo Culture Medium ◽  
Day 3 Embryo

scholarly journals Raman profiling of embryo culture medium to identify aneuploid and euploid embryos

2019 ◽  
Vol 111 (4) ◽  
pp. 753-762.e1 ◽  
Author(s):  
Bo Liang ◽  
Yuan Gao ◽  
Jiabao Xu ◽  
Yizhi Song ◽  
Liming Xuan ◽  
...  
Keyword(s):  
Embryo Culture ◽  
Culture Medium ◽  
Embryo Culture Medium

80 Evaluation of extracellular vesicles from culture medium of human embryos as a possible method of pre-implantation genetic diagnosis

2020 ◽  
Vol 32 (2) ◽  
pp. 166
Author(s):  
C. Aguilera ◽  
D. Veraguas ◽  
C. Henriquez ◽  
A. Velasquez ◽  
F. O. Castro ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Embryo Culture ◽  
Culture Medium ◽  
Chromosomal Abnormalities ◽  
Genetic Diagnosis ◽  
Poor Quality ◽  
Human Embryos ◽  
Acgh Analysis ◽  
Embryo Culture Medium ◽  
Efficiency Of Selection

Noninvasive methods are the clue to increase the efficiency of invitro-derived embryo selection without decreasing their competence. Embryos selection based on their morphology is the most used method but only 40% of selected embryos are able to implant and develop correctly. In humans, pre-implantation genetic diagnosis increases the efficiency of selection by excluding embryos with chromosomal abnormalities. However, pre-implantation genetic diagnosis needs embryonic cells, which might compromise embryo viability. On the other hand, embryos release extracellular vesicles (EVs: microvesicles and exosomes) to the culture medium that contain biological cargo-like proteins and mRNA lipids, and might contain genomic DNA (gDNA). For this study we evaluated the culture medium from embryos generated by intracytoplasmic sperm injection in a certified fertility clinic. Embryos were cultured in Global Total serum-free medium. The embryos were assessed at Day 3 of development and classified in three categories: top, fair, and poor quality. Corresponding medium was collected for isolation of EVs. The nature of EVs was confirmed by their size and concentration using nanoparticle tracking analysis (NTA), presence of surface markers (CD9, CD63, CD81, and CD40L), and morphology using transmission electron microscopy. A correlation analysis between NTA results (EV size and concentration) and embryo quality was performed. To evaluate chromosomal abnormalities of gDNA present in isolated EVs from embryo culture medium, microarray-based comparative genomic hybridization (aCGH) assay was performed. In a second experiment, aCGH analysis was performed and compared between arrested embryos and EVs isolated from corresponding culture medium. Isolated nanoparticles from embryo culture medium were positive to all markers CD9 (30.9%), CD63 (27.2%), CD81 (31.7%), CD40L (8.7%) and had a morphology accordingly to exosomes. The analysis of NTA data indicated that top-quality embryos had EVs with higher diameter (mean: 112.17nm, mode: 91.74nm) than embryos classified as fair (mean: 108.02nm, mode: 89.67nm) and poor quality (mean: 102.78nm, mode: 88.17 nm; P<0.05). The aCGH analysis showed the representation of the 23 pairs of chromosomes in EVs from culture medium and the chromosomal abnormalities were detected in chromosome 4 (C4: 6/15 (40%)) and chromosome 13 (C13: 6/15 (40%)). In the second experiment, the aCGH assay also showed abnormalities in different chromosomes from samples of EVs from culture medium (24.9%) and were more frequent than those observed in the arrested embryos (8.7%; P=0.03). However, the rate of similitude in chromosomal abnormalities between EVs and their respective embryo was 70-80%. In conclusion, the size and gDNA of EVs from culture medium might be an alternative to evaluate the competence of human embryos. This research was supported by FONDECYT-1170310 and Corfo 17Cote-72437, Chile.

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