Development in vitro of preimplantation embryos from 55 mouse strains

Mouse pronuclear embryos were obtained by in vitro fertilization with oocytes of 55 strains of immature females by gonadotrophin-induced ovulation and epididymal sperm of Slc:ICR strain. The number of oocytes ovulated with hormones (5.3-49.4 oocytes per head; average, 22.6), rates of in vitro fertilization (12.2-95.3%; average, 70.7%) and subsequent preimplantation development in Whitten's medium (WM) varied greatly among strains. F1 hybrids developed significantly better than other strains of mice and outbred animals did not develop as well as inbred animals. Within inbred mice, coat colour had no significant effect. In addition, the observation of preimplantation development in WM supplemented with ethylenediamine tetraacetic acid (EDTA) showed that the beneficial effect of the chelator was not universal to mouse embryos; EDTA had a good effect on ICR and PW/aSlc embryos but not on AKR or ddY embryos. The results indicate that strain differences should be considered when interpreting reproductive experiments using mouse embryos.

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A Brief Incubation of Cumulus-Enclosed Mouse Eggs in a Calcium-Free Medium Containing a High Concentration of Calcium-Chelator Markedly Improves Preimplantation Development

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph17103505 ◽

2020 ◽

Vol 17 (10) ◽

pp. 3505

Author(s):

Valeria Merico ◽

Silvia Garagna ◽

Maurizio Zuccotti

Keyword(s):

In Vitro Fertilization ◽

Mammalian Species ◽

Developmental Rate ◽

Cumulus Cells ◽

Fertilization Rate ◽

Preimplantation Development ◽

High Concentration ◽

Vitro Fertilization ◽

Positive Effects

The presence of cumulus cells (CCs) surrounding ovulated eggs is beneficial to in vitro fertilization and preimplantation development outcomes in several mammalian species. In the mouse, this contribution has a negligible effect on the fertilization rate; however, it is not yet clear whether it has positive effects on preimplantation development. Here, we compared the rates of in vitro fertilization and preimplantation development of ovulated B6C3F1 CC-enclosed vs. CC-free eggs, the latter obtained either after a 5 min treatment in M2 medium containing hyaluronidase or after 5–25 min in M2 medium supplemented with 34.2 mM EDTA (M2-EDTA). We found that, although the maintenance of CCs around ovulated eggs does not increment their developmental rate to blastocyst, the quality of the latter is significantly enhanced. Most importantly, for the first time, we describe a further quantitative and qualitative improvement, on preimplantation development, when CC-enclosed eggs are isolated from the oviducts in M2-EDTA and left in this medium for a total of 5 min prior to sperm insemination. Altogether, our results establish an important advancement in mouse IVF procedures that would be now interesting to test on other mammalian species.

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The roles of pyruvate, lactate and glucose during preimplantation development of embryos from F1 hybrid mice in vitro

Development ◽

10.1242/dev.112.1.99 ◽

1991 ◽

Vol 112 (1) ◽

pp. 99-105 ◽

Cited By ~ 2

Author(s):

J.J. Brown ◽

D.G. Whittingham

Keyword(s):

Inbred Mouse ◽

Blastocyst Stage ◽

Preimplantation Development ◽

F1 Hybrids ◽

Mouse Strains ◽

F1 Hybrid ◽

Morula Stage ◽

Lactate And Pyruvate ◽

Hybrid Mice

Embryos of certain inbred mouse strains, and their F1 hybrids, are able to develop from the 1-cell to blastocyst stage in simple chemically defined media containing lactate (L), pyruvate (P) and glucose (G). The individual roles of these substrates in supporting complete preimplantation development in vitro was examined with 1-cell F2 embryos from B6CBF1 hybrid mice. Embryos collected between 26 and 27 h post hCG were cultured in medium containing L, P, LP or LPG. After 50 h in culture, the proportions developing to the morula stage were 1%, 83%, 94% and 100%, respectively. In combination, lactate and pyruvate appeared to act synergistically and both the rate and level of development to the morula stage were unaffected by the absence of glucose. After a further 46 h in culture, only the embryos grown in the presence of glucose developed into blastocysts. In LP medium, embryos arrested at the compacted morula stage late on day 3 of development. As culture continued in the absence of glucose, embryos decompacted (approximately 82 h post hCG) and subsequently degenerated. Exposure to medium containing glucose for the first, second or third 24 h period in culture was sufficient to support the morula-to-blastocyst transition. Glucose still supported this transition when embryos were transferred to LPG medium 3 h after the completion of compaction (76 h post hCG), but was ineffective 6 h later (82 h post hCG) once decompaction had commenced. We conclude that lactate and pyruvate together are able to support normal development of 1-cell F2 embryos to the morula stage in vitro, but that glucose is an essential component of the culture medium for development to the blastocyst stage.

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