Effect of maternal aging and vitrification on mitochondrial DNA copy number in embryos and spent culture medium

Aoki, Sogo; Ito, Jun; Hara, Shunsuke; Shirasuna, Koumei; Iwata, Hisataka

doi:10.1016/j.repbio.2021.100506

Cited by 7 publications

(11 citation statements)

References 56 publications

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“…Four elements which are endogenous retroviruses (which belong in the LTR retrotransposons class) were significantly downregulated in the vitrified oocytes group, but they are not among the most expressed families in human oocytes and the differences were very limited. Vitrification has also been suggested to reduce the copy number of mitochondrial DNA in mice and cows [ 2 , 4 ]. mtDNA is a key determinant in oocyte quality and low levels of mtDNA might indicate low developmental competence [ 34 , 39 ].…”

Section: Discussionmentioning

confidence: 99%

Comparison of oocyte vitrification using a semi-automated or a manual closed system in human siblings: survival and transcriptomic analyses

et al. 2022

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Background Indications of oocyte vitrification increased substantially over the last decades for clinical and ethical reasons. A semi-automated vitrification system was recently developed making each act of vitrification reproducible. In this study, we evaluated the efficiency of the semi-automated technique of oocyte vitrification by survival rate, morphometric assessment and resistance to empty micro-injection gesture as compared with a manual method. Additionally, we intended to evaluate transcriptomic consequences of both techniques using single-cell RNA-seq technology. Results Post-warming survival rate, oocyte surfaces and resistance to empty micro-injection were comparable between semi-automated and manual vitrification groups. Both oocyte vitrification techniques showed limited differences in the resulting transcriptomic profile of sibling oocytes since only 5 differentially expressed genes were identified. Additionally, there was no difference in median transcript integrity number or percentage of mitochondrial DNA between the two groups. However, a total of 108 genes were differentially expressed between fresh and vitrified oocytes (FDR < 0.05) and showed over-represented of genes related to important cellular process. Conclusions Our results provide reassurance about the influence of semi-automation as compared with the manual vitrification method. Concerning oocyte vitrification itself, no tight common transcriptomic signature associated has been observed across studies. Trial registration NCT03570073.

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Section: Discussionmentioning

confidence: 99%

Comparison of oocyte vitrification using a semi-automated or a manual closed system in human siblings: survival and transcriptomic analyses

et al. 2022

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“…The ATP content in individual oocytes were measured using an ATP assay kit (Toyo-Inc., Tokyo, Japan), as previously described [ 30 ]. Luminescence generated with ATP-dependent luciferin-luciferase was measured using a luminometer (Spark 10 M; Tecan Japan Co., Ltd., Kanagawa, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…Luminescence generated with ATP-dependent luciferin-luciferase was measured using a luminometer (Spark 10 M; Tecan Japan Co., Ltd., Kanagawa, Japan). The mitochondrial DNA copy number in oocytes were measured by quantitative PCR using serial dilution of standard as previously described [ 30 ]. DNA of each oocyte was extracted by heating at 55 °C for 30 min and at 98 °C for 10 min in extraction buffer (Tris-HCl, 20 mM containing Nonidet P-40, 0.9% Tween 20, and 0.9% proteinase K, 0.4 mg/mL).…”

Section: Methodsmentioning

confidence: 99%

Beneficial Effect of Polysaccharide Gel Made of Xanthan Gum and Locust Bean Gum on Bovine Oocytes

Hara

Inoue

Aoki

et al. 2023

IJMS

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The present study examined the effect of polysaccharides gels made of xanthan gum and locust bean gum (gel culture system) on oocyte maturation and explored the molecules causing the beneficial effect of the gel culture system. Oocytes and cumulus cells complexes were collected from slaughterhouse-derived ovaries and cultured on a plastic plate or gel. The gel culture system improved the rate of development to the blastocyst stage. The oocytes that matured on the gel contained high lipid contents and F-actin formation, and the resultant 8-cell stage embryos had low DNA methylation levels compared to their plate counterparts. RNA sequencing of the oocytes and embryos revealed the differentially expressed genes between the gel and plate culture systems, and upstream regulator analysis revealed estradiol and TGFB1 as top activated upstream molecules. The medium of the gel culture system contained higher concentrations of estradiol and TGFB1 than that of the plate cultures system. Supplementation of the maturation medium with either estradiol or TGFB1 resulted in high lipid content in oocytes. In addition, TGFB1 improved the developmental ability of the oocytes and increased F-actin content while reducing DNA methylation levels in the 8-cell stage embryos. In conclusion, the gel culture system is useful for embryo production, potentially through the upregulation of TGFB1.

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“…Moreover, the level of SIRT1 upregulation in response to induced mitochondrial dysfunction was low in oocytes derived from aged cows [12]. Mitochondria damaged by vitrification were actively removed from the embryos of young cows, whereas damaged mitochondrial removal activity was low in aged cows [13]. These findings suggest that certain age-associated events hinder SIRT1 and MQC activation in embryos.…”

Section: Introductionmentioning

confidence: 99%

Age-related advanced glycation end-product accumulation impairs mitochondrial regulation after vitrification

Ito

Iwata

2023

Biology of Reproduction

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Vitrification is an important assisted reproductive technology, although it induces mitochondrial dysfunction in embryos. Herein, we aimed to investigate whether age-associated accumulation of advanced glycation end-products (AGEs) in oocytes impairs the recovery of embryos from cryopreservation-induced mitochondrial dysfunction/damage. Mouse 8-cell stage embryos developed in vitro were vitrified and warmed and incubated up to the blastocyst stage. AGE levels in oocytes were higher in both aged mice and AGE accumulation mouse models (MGO-mice) than those in young and control mice. Additionally, the level of SIRT1 upregulation was lower for embryos of aged and MGO-mice than that for embryos of young and control mice. The highest mtDNA content was detected in blastocysts derived from vitrified embryos of aged and MGO-mice. The spent culture medium of blastocysts derived from both aged and MGO-mice contained higher mtDNA content than that of the blastocysts derived from young and control mice. EX527 increased mtDNA content in the spent culture medium of vitrified embryos derived from young mice. In addition, p62 aggregate levels were higher in vitrified embryos of control mice than those in vitrified embryos of MGO-mice. The SIRT1 activator, resveratrol, increased p62 aggregation levels in vitrified embryos derived from young and aged mice, whereas vitrification did not affect p62 aggregation levels in embryos from aged mice. Therefore, age-associated AGE accumulation induces decreased responsive SIRT1 upregulation following vitrified–warmed treatment and impairs mitochondrial quality control activity in vitrified embryos.

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Effect of maternal aging and vitrification on mitochondrial DNA copy number in embryos and spent culture medium

Cited by 7 publications

References 56 publications

Comparison of oocyte vitrification using a semi-automated or a manual closed system in human siblings: survival and transcriptomic analyses

Comparison of oocyte vitrification using a semi-automated or a manual closed system in human siblings: survival and transcriptomic analyses

Beneficial Effect of Polysaccharide Gel Made of Xanthan Gum and Locust Bean Gum on Bovine Oocytes

Age-related advanced glycation end-product accumulation impairs mitochondrial regulation after vitrification

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