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New Frontiers in IVF: mtDNA and autologous germline mitochondrial energy transfer
Ovarian aging leads to a decrease in the quantity and quality of oocytes and aged oocytes have a reduced number of mitochondria. Mitochondria are the energy factories of the cells and their lacked could leads to lower fertilization rates and poor embryonic development. Various strategies have been tested to increase the mitochondria quantity and thus improve the quality of oocytes used in in vitro fertilization. Results of ovarian rejuvenation te
Conclusions. HPV may have an impact in spermatogenesis, because an HPV infection was more frequently detected in men with abnormal sperm parameters. High-risk HPV 52 was the most common genotype among couples undergoing IVF treatment.
Parameters of the Mouse Embryo Assay that affect detection of peroxides in mineral oil
Conclusions: Current quality control protocols may not be sensitive enough to identify low concentrations of peroxides, which, if undetected, can increase over time and become potentially harmful during gamete and embryo culture. The different parameters established in this study allow the sensitivity of the Mouse Embryo Assays to be optimized to specifically detect peroxides in mineral oil samples prior to their release into the market and their
Epigenetic Changes and Assisted Reproductive Technologies
The reason for this focus is that the developmental time points at which ART clinical and laboratory procedures are implemented are precisely the time points at which large-scale reorganization of the epigenome takes place during normal development. In this manuscript, we review the many human studies comparing the epigenomes of ART children with children conceived in vivo, as well as assess the potential of individual ART clinical and laboratory
Sperm chromosomal abnormalities and their contribution to human embryo aneuploidy
In conclusion, sperm FISH analysis offers clinical prognostic value to evaluate reproductive possibilities in infertile couples. Therefore, couples with abnormal sperm FISH results should be offered genetic counseling and presented with clinical options such as PGT-A.
Association between the number of retrieved mature donor oocytes and live birth in IVF donor recipient cycles using frozen donor eggs [2017 Abstract]
Conclusions: This current analysis of IVF outcomes among recipients of vitrified donor oocytes demonstrates that live birth from fresh embryo transfer is not adversely affected by retrieval of more oocytes. Secondarily, there were more surplus embryos for cryopreservation with increasing numbers of retrieved oocytes, increasing the potential cumulative number of infants per retrieval. These findings must be considered in the context of achieving
Growth Hormone and Reproduction: Lessons Learned From Animal Models and Clinical Trials
Evidence suggests that GH can stimulate the secretion of IGF1 not only in the liver but also in the peripheral target structures, including the ovary. Moreover, IGF-1 can be secreted locally under the influence of stimuli other than GH. In the case of the ovary, steroid hormones, gonadotropins or the combination of both seems to be involved. Even more interesting, the ovary itself can secret GH locally and exert a paracrine action modulating the
Factors influencing dynamics of follicular pool development
Published in Frontiers in Endocrinology (2019).Growth Hormone and Reproduction: Lessons Learned From Animal Models and Clinical Trials. Dosouto C, Calaf J, Polo A, Haahr T, Humaidan P. Front Endocrinol (Lausanne). 2019 Jun 26:10:404. doi: 10.3389/fendo.2019.00404.
Can the sensitivity of the Mouse embryo Assay (MEA) be improved to detect low dose mineral oil toxicity?
Summary
The MEA has been shown to improve its sensitivity by changing the protein supplementation, embryo density and culture dish.
Comment
One of the reasons why all IVF clinics have outsourced their medium preparation for IVF to specialised companies is the problem of toxicity testing. The standard format used by all IVF media companies (and demanded by clinics and accreditation bodies) is the Mouse Embryo Assay (MEA). The MEA tests for the ability of media to support day one mouse embryos development to day 4 (or later) blastocysts. The general minimum standard is a rate for more than 80% blastocysts recovery. Embryologist would largely acknowledge that this assay is the very basic test and that low levels of toxins may remain undetected. The spanish authors in this subscription-based article in RBMO have explored if the standard test could be modified to enhance its sensitivity. The toxin they used was mineral oil that had been infused with a known concentration of peroxide and they tested its effect on mouse blastulation rates by changing the type and concentration of protein and several culture conditions namely embryo density and the type of culture vessel.|Firstly and reassuringly, all but the lowest dose of peroxide were toxic in this test which shows that the test can detect subtly compromised mineral oil. Using a very low dose of peroxide, they essentially reported that reduced development occurred with HSA with or without low dose globulins compared to HSA+high dose globulins and BSA (included as a reference even though it is not used in IVF culture medium). Extending the duration of culture/exposure from 4 to 5days days also improved sensitivity (I.e. less development). Using the same protein testing regimen, they further showed that low number group culture (3ul/embryo) and microwell dishes (25ul) were less sensitive to toxicity than large group culture (25ul/embryo) and petri dishes (50ul droplets).|In summary then, this paper has shown that the standard MEA test has potential to be improved in their detection of low level toxicity beyond what is now offered by media producers with little additional effort by using HAS +/- low dose globulins, restricting the embryo density and prolonging culture times.|By extension, knowing the sensitivity of the test to low dose peroxide doses, one may also run a standard dose response test with a defined LD50 dose where one may expect a certain decrease in development. Any further decrease in development may suggest the presence of other toxins at undetectable limits. It was interesting that in this paper, the authors used a concentration at the lower limit of detection but has some degree of toxicity. It goes to illustrate the sensitivity of the embryo culture system. |One aspect of the paper was that while HSA+/- low dose globulins improved the sensitivity of the assay, the counter argument implies that if toxins exist in the mineral oil (or elsewhere), the HSA protein supplementation will facilitate it transfer into the embryo over and above BSA or high dose HSA (+low dose globulins). The authors noted this and suggested that HSA may be more efficient in the transport of nutrients into the embryo. Using a more sensitive approach should mean that media, including mineral oil, has less chance of containing low level toxicity and overall should mean better IVF outcomes. |However, it does imply that HSA on its own may be less suitable if mineral oil toxicity develops after the initial testing. After all, there is some time between testing and final use of a product. Give that many clinics purchase their oil in large volumes and the same bottle is accessed multiple times, a clinic manger cannot be sure that the oil has not degraded over time with multiple exposures to the atmosphere. One option is to purchase oil in lower volume bottles and limit their multiple assess. Purchasing oil that has been tested with hyper sensitive procedures is another approach as is repeated testing over the life of the media or oil. Other authors have suggested time-lapse check points are an alternative way to improve sensitivity. |Finally, many of the tools the authors suggest may improve sensitivity are also the changes that are now dominating embryo culture. In particular, the lower embryo density and droplet volume are part of a current progression into time-lapse monitoring and individual tracking of embryos. The reason for raising this is that these changes have the potential to increase the risk to embryo development if for whatever reason, the quality of the environment decreases. Knowing this is one of the risks of using smaller droplets and individual culture, quality managers should ensure that monitoring is in place, as far as practicable, to detect subtle changes in the environment. After all, even though the media may pass MEA testing, does mean that every dish or pipette or gas bottle are ‘pure’. Knowing that individual embryo culture may deliver better outcomes and at the same time makes them more sensitive to potential stress is half the battle.|IVFCPD thanks Dr Mestres for checking the accuracy of this commentary.|
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In this test, an improved assay sensitivity resulted in:
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With respsect to embryo density, the authors reported improved sensitivity when
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With respect to culture volume, the authors reported improved sensitivity when
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With respect to protein type and concentration, the authors reported some protein and doses improved the sensitivity of the MEA. Which of the following were reported to result in fewer blastocysts when exposed to toxic mineral oil compared to the BSA.
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Vit Kit - Warm NX is an adaptable, cost-effective system for use in the thawing of oocytes, pronuclear zygotes, cleavage stage embryos, and blastocyst stage embryos. Unlike many vitrification kits that feature a mono-buffered system and M199 base med
Vit Kit - Freeze NX is an adaptable, cost-effective system for use in the vitrification of oocytes, pronuclear zygotes, cleavage stage embryos, and blastocyst stage embryos. Vit Kit - Freeze NX is the latest advancement in vitrification media aimed t
S-Cryolock is the slimmer version of the original. It is a versatile, simple and efficient vitrification device that is intended for the holding, cryopreservation and storage of oocytes or embryos in liquid nitrogen.
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