Washing mineral oil reduces contaminants and embryotoxicity

Morbeck, Dean E.; Khan, Zaraq; Barnidge, David R.; Walker, David

doi:10.1016/j.fertnstert.2010.03.067

Cited by 54 publications

(50 citation statements)

References 15 publications

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“…However, this is not the only potential contaminant that may be present in products used for embryo culture. A variety of other chemical contaminants have been detected in different culture supplies, including peroxides, alkenes, and octanoic acid [2,3,41]. Similarly, some components of the Our results indicate that a MEA utilizing IVM-derived embryos from SW mice is highly sensitive to TX100, since multiple endpoints detected low levels of this contaminant.…”

Section: Discussionmentioning

confidence: 50%

“…TX100 was chosen as a representative contaminant, since this chemical has been detected in culture supplies and used in other studies evaluating the suitability of MEA conditions [2,8,40]. In addition, two studies have shown that the effects of TX100 are similar to those of two other known contaminants [8,40].…”

Section: Discussionmentioning

confidence: 99%

“…Even when the highest-quality reagents and plasticware are used, the introduction of some unknown substances (contaminants) into the culture environment is unavoidable [1][2][3]. As a result, all supplies that will come into contact with the embryo and/or the culture medium should be subjected to strict quality control (QC) testing, including the ability to support the growth of murine embryos [4,5].…”

Section: Introductionmentioning

confidence: 99%

“…Using two low concentrations of a known contaminant of culture products (Triton X-100) [2,8], the objective of this series of experiments was to compare the sensitivities of one-cell embryos from hybrid mice and embryos produced by IVM and IVF from hybrid mice, as well as 3 strains of outbred mice using multiple developmental parameters (morphological development, blastocyst cell number and allocation, and blastocyst gene expression). [26] contained 0.5 mM glucose, 0.2 mM pyruvate, 6.0 mM lactate, 0.5 mg/mL insulin, 0.275 mg/mL transferrin, 0.25 ng/mL selenium, 10 ng/mL epidermal growth factor (EGF), 2 mg/mL fetuin, and 2.5 mg/ mL recombinant human albumin (AlbIX; Novozymes, Bagsvaerd, Denmark).…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 50%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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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“…First, in addition to peroxide, multiple compounds may contribute to mineral oil toxicity [22,23] and include malonaldehyde and alkenals as products of peroxide activity. Thus, the effects observed cannot be ascribed only to peroxide.…”

Section: Discussionmentioning

confidence: 99%

Improved detection of mineral oil toxicity using an extended mouse embryo assay

Ainsworth

Fredrickson

Morbeck

2017

J Assist Reprod Genet

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Purpose Successful in vitro fertilization (IVF) relies on sound laboratory methods and culture conditions which depend on sensitive quality control (QC) testing. This study aimed to improve the sensitivity of mouse embryo assays (MEA) for detection of mineral oil toxicity. Methods Five experiments were conducted to study modifications of the standard mouse embryo assay (MEA) in order to improve sensitivity using clinical grade mineral oil with known peroxide concentrations. Assessment of blastocyst development at either 96 h or in an extended MEA (eMEA) to 144 h was tested in each experiment. In experiment 1, ability to detect peroxides in oil was compared in the MEA, eMEA, and cell number at 96 h. In experiment 2, serial dilutions of peroxide in oil were used along with time-lapse imaging to compare sensitivity of the morphokinetic MEA to the eMEA. Culture conditions that may affect assay sensitivity were assessed in experiments 3-5, which examined the effect of group versus individual culture, oxygen concentration, and protein supplementation. Results Extended MEA and cell counts identified toxicity not detected by the routine endpoint of blastocyst rate at 96 h. The eMEA was fourfold more sensitive than the standard MEA, and this sensitivity was similar to the morphokinetic MEA. Group culture had a protective effect against toxicity, while oxygen concentration did not affect blastocyst development. Protein supplementation with HSA had a protective effect on blastocyst development in eMEA.Conclusions The standard MEA used by manufacturers does not detect potentially lethal toxicity of peroxides in mineral oil. While group culture may mask toxicity, protein supplementation and oxygen concentration have minimal effect on assay sensitivity. The eMEA and time-lapse morphokinetic assessment are equally effective in detection of peroxide toxicity and thus provide manufacturers and end-users a simple process modification that can be readily adopted into an existing QC program.

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Production of Multiple Cell‐Laden Microtissue Spheroids with a Biomimetic Hepatic‐Lobule‐Like Structure

Gao

Kim

et al. 2021

Advanced Materials

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The construction of an in vitro 3D cellular model to mimic the human liver is highly desired for drug discovery and clinical applications, such as patient‐specific treatment and cell‐based therapy in regenerative medicine. However, current bioprinting strategies are limited in their ability to generate multiple cell‐laden microtissues with biomimetic structures. This study presents a method for producing hepatic‐lobule‐like microtissue spheroids using a bioprinting system incorporating a precursor cartridge and microfluidic emulsification system. The multiple cell‐laden microtissue spheroids can be successfully generated at a speed of approximately 45 spheroids min−1 and with a uniform diameter. Hepatic and endothelial cells are patterned in a microtissue spheroid with the biomimetic structure of a liver lobule. The spheroids allow long‐term culture with high cell viability, and the structural integrity is maintained longer than that of non‐structured spheroids. Furthermore, structured spheroids show high MRP2, albumin, and CD31 expression levels. In addition, the in vivo study reveals that structured microtissue spheroids are stably engrafted. These results demonstrate that the method provides a valuable 3D structured microtissue spheroid model with lobule‐like constructs and liver functions.

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Washing mineral oil reduces contaminants and embryotoxicity

Cited by 54 publications

References 15 publications

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

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

Improved detection of mineral oil toxicity using an extended mouse embryo assay

Production of Multiple Cell‐Laden Microtissue Spheroids with a Biomimetic Hepatic‐Lobule‐Like Structure

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