Peroxidation of mineral oil used in droplet culture is detrimental to fertilization and embryo development

Otsuki, Junko; Nagai, Yasushi; Chiba, Kazuyoshi

doi:10.1016/j.fertnstert.2006.11.144

Cited by 76 publications

(75 citation statements)

References 2 publications

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“…Mineral oil used for IVF has been shown to undergo peroxidation under certain storage conditions [3]. In the present study, cumene hydroperoxide was used as a model for naturally occurring peroxides in mineral oil.…”

Section: Discussionmentioning

confidence: 99%

“…Though oil is important for pH, temperature and osmolar stability, it is a petroleum product that can harbor embryo toxins, including zinc [2] and peroxides [3]. Recently, Otsuki and colleagues reported peroxide contamination of laboratory grade mineral oil and found the degree of peroxidation was dependent on exposure to heat, UV light, extended storage, manufacturer and lot number [3,4]. Peroxide-contaminated oil was also shown to adversely affect murine embryo development and in vitro fertilization success [3].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Peroxides in mineral oil used for in vitro fertilization: defining limits of standard quality control assays

Hughes¹,

Morbeck

Hudson³

et al. 2010

J Assist Reprod Genet

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Purpose To determine the relative sensitivities of the 1 and 2-cell mouse embryo assays (MEA) and the human sperm motility assay (HSMA) for peroxides in mineral oil. The effect of peroxide on blastocyst cell number and apoptosis was also studied. Methods One and two-cell MEA and HSMA were performed using mineral oil containing cumene hydroperoxide (CH).Results The 1-cell MEA was twice as sensitive as the 2-cell MEA and 20-times more sensitive than the HSMA for CH in mineral oil. The sensitivity of the 1-cell MEA doubled when embryos were cultured individually versus group culture. CH decreased blastocyst cell number in a dose dependent manner.Conclusions Individually cultured 1-cell embryos had the highest sensitivity for peroxides in mineral oil. Current quality control assays, including group cultured murine embryos and human sperm motility, have limited sensitivity for peroxides in mineral oil and may not detect levels of peroxides that cause sub-lethal cellular damage.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Peroxides in mineral oil used for in vitro fertilization: defining limits of standard quality control assays

Hughes¹,

Morbeck

Hudson³

et al. 2010

J Assist Reprod Genet

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“…Mineral oil, a standard component of embryo culture systems that is poorly defined, variable from batch to batch, and has the capacity to become toxic over time, remains the IVF laboratory's product with the most potential for variation in quality [7]. While previous data indicate that toxicity of manufacturer recalled oil can be detected with improved MEA methods [5,6], neither method was suitable for routine use in clinical laboratories.…”

Section: Discussionmentioning

confidence: 99%

“…Derived from crude oil, mineral oil contains aromatic and unsaturated hydrocarbons [7,8] that are susceptible to peroxidation and free radical formation, known embryo toxins that can develop during storage and thus adversely affect embryo development and IVF outcomes even after passing testing by the manufacturer [7,9,10]. The chemical nature of mineral oil makes it the product in the IVF lab with the highest risk for adversely affecting clinical outcomes and warrants additional, end-user scrutiny.…”

Section: Introductionmentioning

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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“…Cumene hydroperoxide (CH; Sigma-Aldrich) was used as a surrogate for peroxides that can accumulate naturally in mineral oil (Otsuki et al, 2007;Morbeck et al, 2010) and was prepared as previously described (Hughes et al, 2010). For each replicate (n ¼ 3), an EmbyoSlide (n ¼ 5/replicate) was prepared for each concentration of CH (0, 2, 4, 6 or 8 mM).…”

Section: Experiments One: Cumene Hydroperoxidementioning

confidence: 99%

Advances in quality control: mouse embryo morphokinetics are sensitive markers of in vitro stress

Wolff

Fredrickson

Walker

et al. 2012

Fertility and Sterility

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what is known already: Current quality control testing methodology is sensitive but further improvements are needed to assure optimal culture conditions. MKs of embryo development may detect small variations in culture conditions. study design: Cross sectional-control versus treatment. Mouse embryo development kinetics of 466 embryos were analyzed according to exposure to various concentrations of toxins and toxic mineral oil. materials, setting, methods: Cryopreserved 1-cell embryos from F1 hybrid mice were cultured with cumene hydroperoxide (CH) (0, 2, 4, 6 and 8 mM) and Triton X-100 (TX-100; 0, 0.0008, 0.0012, 0.0016 and 0.002%). Using the Embryoscope, time-lapse images were obtained every 20 min for 120 h in seven focal planes. End-points were timing and pattern of cell division and embryo development. The blastocyst rate (BR) was defined as the percentage of embryos that developed to the expanded blastocyst stage within 96 h. main results and the role of chance: BR was not affected for embryos cultured in the three lowest concentrations of CH and the four lowest concentrations of TX-100. In contrast, a unique MK model detected all concentrations tested (P , 0.05). The MK model identified toxicity in two lots of toxic mineral oil that did not affect BR (P , 0.05).limitations, reasons for caution: A limited number of toxins were used so that the results may not apply to all potential embryo toxins. A larger sample size may also demonstrate other statistically significant developmental kinetic parameters.wider implications of the findings: MKs in mouse embryos are a sensitive and efficient method for quality control testing of in vitro culture conditions. BR, the end-point of traditional quality control assays, did not detect sublethal concentrations of toxins in the culture milieu in our study. This study demonstrates that temporal variation at key developmental stages reflects the quality of the culture environment. An MEA that incorporates MK will provide enhanced sensitivity and faster turn-around times.

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Peroxidation of mineral oil used in droplet culture is detrimental to fertilization and embryo development

Cited by 76 publications

References 2 publications

Peroxides in mineral oil used for in vitro fertilization: defining limits of standard quality control assays

Peroxides in mineral oil used for in vitro fertilization: defining limits of standard quality control assays

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

Advances in quality control: mouse embryo morphokinetics are sensitive markers of in vitro stress

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