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. 2020 Mar 13;102(3):521-523.
doi: 10.1093/biolre/ioz224.

Oocytes from female mice on MF1 genetic background are not suitable for assisted reproduction†

Yasushiro Yamauchi  1 , Anna Ajduk  1   2 , Monika A Ward  1
Affiliations

Oocytes from female mice on MF1 genetic background are not suitable for assisted reproduction†

Yasushiro Yamauchi et al. Biol Reprod. .
No abstract available

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Figures

Figure 1
Figure 1
Oocytes from female mice on MF1 genetic background are not suitable for assisted reproduction. (A) Exemplary images of embryos produced after in vitro fertilization with gametes from MF1 mice. Metaphase II (MII) oocytes retrieved from MF1 females (i) were in vitro fertilized by sperm from MF1 males. Formation of fertilization cone (*) occurred normally (ii), followed by normal second polar body extrusion and formation of pronuclei. However, as the pronuclei stage progressed, the zygotes began to lose their smooth oval shape (iii, v, arrowheads). The deformity was the most apparent at the two-cell stage, with shape of blastomeres highly irregular and blebbing (arrowheads) (iv, vi). Scale = 50 μm. (B) Comparison of developmental potential of embryos produced with oocytes from MF1 and B6D2F1 females. (C) Summary of ART modifications attempted in order to overcome embryo deformity. (D) In vitro development of in vivo and in vitro generated embryos. Zygotes were obtained after in vitro (i–iii) and in vivo (iv) fertilization and were cultured for 96 h either in CZB (i, iii, iv) or KSOM (ii–iii) culture medium. The percentage of embryos was calculated from oocytes inseminated (i–ii), embryos of a given stage (iii), and activated oocytes retrieved from female reproductive track after mating (iv). The number of oocytes inseminated/retrieved is shown in parentheses under the X axis (i–iii) and the numbers from which percentage was derived are shown above the graphs (iii). In crosses, a female is shown first. The data in i-iii are shown as mean ± SEM, with n = 3 independent experiments. The data in iv represent a single experiment. Raw data are shown in Supplementary Table S1. Statistical significance (Fisher’s exact test, P < 0.05): asignificantly different than respective developmental stage in B6D2F1 × MF1; bsignificantly different than respective developmental stage in the same cross cultured in KSOM (shown in panel ii). (E) Effect of oxygen tension on in vitro generated MF1 embryos. Zygotes were obtained after in vitro fertilization and were cultured in CZB medium under two different oxygen tensions (5,5: 5% CO2, 5% O2; 5,21: 5% CO2, 21% O2), and were either transferred at the two-cell stage (i) or cultured for 96 h (ii). The percentages of deformed zygotes and deformed two-cell embryos were calculated from zygotes and two-cell embryos obtained, respectively. The percentages of live offspring and blastocysts were calculated from two-cell transferred or cultured, respectively. The numbers from which percentages were derived are shown above the bars. The data in i and ii are derived from two and three independent experiments, respectively. Raw data are shown in Supplementary Tables S2 and S3. Statistical significance (Fisher’s exact test): different than respective category in 5,5: **P < 0.01; ****P < 0.0001. (F) Pronuclear transfer. B6D2F1 (F1) and MF1 MII oocytes were chemically activated. Between 4 and 5 h post-activation, pronuclei (MF1 and F1 nuclei) were removed and transplanted reciprocally to enucleated MF1 or F1 oocytes (MF1 cyto or F1 cyto). F1 nuclei transplanted to F1 cyto served as a control for surgery effects and activated unmanipulated MF1 and F1 oocytes served as positive and negative control for deformity. Twenty-four hours after activation embryos were scored as normal, mildly deformed (one or more cytofragments smaller in size than the size of polar body and/or evident blebbings), and severely deformed (more small-sized fragments or fragments of larger size with total volume of fragments comprising an approximately equivalent of one-half of one blastomere of greater). The data represent pooled data from two independent experiments. The numbers above the graphs show the number of oocytes from which percentages were derived.
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References

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