Potential of inner cell mass outgrowth and amino acid turnover as markers of quality in the in vitro fertilization laboratory

Gada, R.; Daftary, Gaurang S.; Walker, David; Lacey, Jean M.; Matern, Dietrich; Morbeck, Dean E.

doi:10.1016/j.fertnstert.2012.06.012

Cited by 12 publications

(13 citation statements)

References 31 publications

(41 reference statements)

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“…Embryos were cultured for 3 days, and then blastocysts (4.5 dpc) were transferred into a fibronectin (FN)‐coated organ dish (Sigma). Thereafter they were cultured to generate outgrowth embryos in Dulbecco's modified Eagle's media (Gibco/Life Technologies, Carlsbad, CA) without any supplementation for 3 days (7.5 dpc; Gada et al, ). Overall 300 μl of outgrowth embryo–conditioned medium (OECM) was collected from 150 outgrowth embryos (2 μl/outgrowth embryo) and transferred to an Eppendorf tube, and stored at −80°C until EVs isolation.…”

Section: Methodsmentioning

confidence: 99%

Embryotrophic effects of extracellular vesicles derived from outgrowth embryos in pre‐ and peri‐implantation embryonic development in mice

Kim

Lee

et al. 2018

Molecular Reproduction Devel

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Recently, many studies have investigated the role of extracellular vesicles (EVs) on reproductive events, including embryo development and death, oviduct–embryo crosstalk, in vitro fertilization and others. The aim of this study was to demonstrate whether outgrowth embryo–derived EVs function as bioactive molecules and regulate mouse embryonic developmental competence in vitro and implantation potential in utero. The EVs from mouse outgrowth embryos on 7.5 days postcoitum were detected and selectively isolated to evaluate the embryotrophic functions on preimplantation embryos. Developmental outcomes such as the percentage of blastocyst formation, hatching, and trophoblastic outgrowth were assessed. Furthermore, the total cell number and apoptotic index of blastocysts, which were incubated with EVs during the culture period, were evaluated by fluorescence microscopy. Implantation potential in utero was investigated following embryo transfer. The EVs from outgrowth embryo–conditioned media have rounded membrane structures that range in diameter from 20 to 225 nm. Incubation with EVs improved preimplantation embryonic development by increasing cell proliferation and decreasing apoptosis in blastocysts. Moreover, the implantation rates following embryo transfer were significantly higher in EV–supplemented embryos compared with the control. Collectively, EVs from outgrowth embryo could enhance the embryonic developmental competence and even implantation potential in mice.

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Section: Methodsmentioning

confidence: 99%

Embryotrophic effects of extracellular vesicles derived from outgrowth embryos in pre‐ and peri‐implantation embryonic development in mice

Kim

Lee

et al. 2018

Molecular Reproduction Devel

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“…Most MEAs simply determine the proportion of embryos that have reached the blastocyst stage by the end of the culture period (blastocyst quantity), without addressing the possibility of inhibitory effects on blastocyst quality. Total cell number of the blastocyst, the number of cells in the trophectoderm (TE) and inner cell mass (ICM), outgrowth of hatched blastocysts, and embryo metabolism can all be affected by culture conditions, providing alternative endpoints for QC testing that may result in increased sensitivity [8,14,[19][20][21]. For example, ammonium in the culture medium can alter cell number, apoptosis, cellular metabolism, gene expression, and fetal development with no effect on blastocyst development [22,23].…”

Section: Introductionmentioning

confidence: 99%

Building a better mouse embryo assay: effects of mouse strain and in vitro maturation on sensitivity to contaminants of the culture environment

Herrick

Paik

Strauss

et al. 2015

J Assist Reprod Genet

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Purpose The aim of this study is to compare the sensitivity of the standard one-cell mouse embryo assay (MEA) to that using in vitro-matured oocytes from hybrid and outbred mice. Methods The study was done by culturing embryos in the presence or absence of two concentrations (0.0005 or 0.001 %v/v) of Triton X-100 (TX100). Embryonic development, blastocyst cell numbers (total and allocation to the trophectoderm [TE] and inner cell mass [ICM]), and blastocyst gene expression were evaluated. Results Neither concentration of TX100 affected (P>0.05) cleavage, blastocyst development, or hatching in one-cell embryos from BDF1 mice. However, all cell number endpoints were reduced (P<0.05) by the high concentration of TX100 and the number of ICM cells was reduced (P<0.05) by the low concentration of TX100. Inhibitory (P<0.05) effects of the high concentration of TX100 were observed in in vitro maturation (IVM) embryos from BDF1, CF1, and SW, but not ICR, mice. Cell number and allocation were negatively affected by the high concentration of TX100 in CF1 and SW embryos, but not in BDF1 or ICR embryos. The only developmental endpoints affected by the low concentration of TX100 were cleavage of BDF1 oocytes, blastocyst development of SW embryos, and cell numbers (total and inner cell mass (ICM)) of SW blastocysts. Conclusions The sensitivity of the MEA to TX100 is improved by using embryos from in vitro-matured oocytes, using oocytes from some outbred (SW or CF1, not ICR) strains of mice, and evaluating blastocyst cell number and allocation.

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“…This would not only significantly reduce the chance that these products would reach the ART laboratory but would also decrease resources used for testing. Metabolomics can similarly be used to detect changes in metabolism caused by low levels of embryo-toxic substances (65).…”

Section: Applications Of Metabolomic Technology To Artmentioning

confidence: 99%

Omics as a window to view embryo viability

Krisher

Schoolcraft

Katz‐Jaffe

2015

Fertility and Sterility

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Potential of inner cell mass outgrowth and amino acid turnover as markers of quality in the in vitro fertilization laboratory

Cited by 12 publications

References 31 publications

Embryotrophic effects of extracellular vesicles derived from outgrowth embryos in pre‐ and peri‐implantation embryonic development in mice

Embryotrophic effects of extracellular vesicles derived from outgrowth embryos in pre‐ and peri‐implantation embryonic development in mice

Building a better mouse embryo assay: effects of mouse strain and in vitro maturation on sensitivity to contaminants of the culture environment

Omics as a window to view embryo viability

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