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Advances in quality control: mouse embryo morphokinetics are sensitive markers of in vitro stress
H S Wolff1, J R Fredrickson, D L Walker
1Department of Obstetrics and Gynecology, Mayo Clinic, Rochester, MN 55905, USA.
Study Question:
Can time-lapse analysis of cell division timings [morphokinetics (MK)] in mouse embryos detect toxins at concentrations that do not affect blastocyst formation?
Summary Answer:
An MK algorithm enhances assay sensitivity while providing results 24-48 h sooner than the traditional mouse embryo assay (MEA).
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 µM) 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.
Insights
Morphokinetics (MK) analysis of mouse embryos detects toxins missed by traditional blastocyst rate (BR) assays. This enhanced method offers greater sensitivity and faster results for in vitro culture quality control.
Area of Science:
- Embryology
- Toxicology
- Reproductive Biology
Background:
- Current quality control methods for in vitro culture are sensitive but require improvement.
- Embryo morphokinetics (MK) may reveal subtle variations in culture conditions.
- Time-lapse imaging offers potential for enhanced embryo assessment.
Purpose of the Study:
- To determine if time-lapse analysis of mouse embryo morphokinetics (MK) can detect toxins at concentrations not affecting blastocyst formation (BR).
- To evaluate the sensitivity of MK compared to traditional assays for detecting toxicity in culture media.
- To assess the potential of MK for improving quality control in assisted reproductive technologies.
Main Methods:
- Analysis of 466 mouse embryos using time-lapse imaging (Embryoscope) over 120 hours.
- Exposure of embryos to varying concentrations of cumene hydroperoxide (CH) and Triton X-100 (TX-100).
- Assessment of cell division timings, patterns, and overall embryo development, including blastocyst rate (BR).
Main Results:
- Blastocyst rate (BR) was unaffected by low concentrations of CH and TX-100.
- A morphokinetics (MK) algorithm detected toxicity at all tested concentrations (P < 0.05).
- MK identified toxicity in mineral oil lots that did not impact BR (P < 0.05).
Conclusions:
- Morphokinetics (MK) in mouse embryos provides a sensitive and efficient method for in vitro culture quality control.
- MK analysis detects sublethal toxin concentrations missed by traditional blastocyst rate (BR) assays.
- Incorporating MK into embryo assays enhances sensitivity and reduces turnaround time compared to traditional methods.
