Quantitative Analysis of Morphological Modifications of Day 6.5 Horse Embryos after Treatment with Four Cryoprotectants: Differential Effects on Inner Cell Mass and Trophoblast Cells1

Bruyas, J.F.; Battut, I.; Pol, J-M.; Botrel, C.; Fieni, F.; Tainturier, D.

doi:10.1093/biolreprod/52.monograph_series1.329

Cited by 14 publications

(26 citation statements)

References 46 publications

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“…2). This has been demonstrated previously not only in in vitro-produced equine embryos (Tremoleda et al 2003) but also in early equine blastocysts that were recovered from mares on day 6 after ovulation (Bruyas et al 1993(Bruyas et al , 1995. We found that the central cell network of the horse embryo consists of a mixture of EPI and PE cells, which is similar to findings in very early blastocysts in mice and cattle (Chazaud et al 2006, Kuijk et al 2012, Schrode et al 2014.…”

Section: Treatmentsupporting

confidence: 88%

Cell lineage allocation in equine blastocysts produced in vitro under varying glucose concentrations

et al. 2015

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Equine embryos develop in vitro in the presence of high glucose concentrations, but little is known about their requirements for development. We evaluated the effect of glucose concentrations in medium on blastocyst development after ICSI. In experiment 1, there were no significant differences in rates of blastocyst formation among embryos cultured in our standard medium (DMEM/F-12), which contained O16 mM glucose, and those cultured in a minimal-glucose embryo culture medium (!1 mM; Global medium, GB), with either 0 added glucose for the first 5 days, then 20 mM (0-20) or 20 mM for the entire culture period . In experiment 2, there were no significant differences in the rates of blastocyst development (31-46%) for embryos cultured in four glucose treatments in GB (0-10, 0-20, 5-10, or 5-20). Blastocysts were evaluated by immunofluorescence for lineage-specific markers. All cells stained positively for POU5F1. An inner cluster of cells was identified that included presumptive primitive endoderm cells (GATA6-positive) and presumptive epiblast (EPI) cells. The 5-20 treatment resulted in a significantly lower number of presumptive EPI-lineage cells than the 0-20 treatment did. GATA6-positive cells appeared to be allocated to the primitive endoderm independent of the formation of an inner cell mass, as was previously hypothesized for equine embryos. These data demonstrate that equine blastocyst development is not dependent on high glucose concentrations during early culture; rather, environmental glucose may affect cell allocation. They also present the first analysis of cell lineage allocation in in vitro-fertilized equine blastocysts. These findings expand our understanding of the factors that affect embryo development in the horse.

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Section: Treatmentsupporting

confidence: 88%

Cell lineage allocation in equine blastocysts produced in vitro under varying glucose concentrations

et al. 2015

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“…The ‘classical’ CPA for slow‐freezing horse embryos is glycerol (10%: 1.36 M) with which the embryo is equilibrated by incubation in 2–4 solutions of increasing concentration. Other CPAs, such as DMSO, 1,2 propanediol, ethylene glycol (1.5 M; Bruyas et al. 1995) and methanol (2.5 M: Bass et al.…”

Section: Freezing Techniquesmentioning

confidence: 99%

“…2004), have also been tested (for an extensive review of CPA concentrations and times of exposure tested; see Bruyas 2011), primarily because early studies suggested that exposure to glycerol, rather than freezing per se , was responsible for most of the damage suffered by embryos at the level of cellular ultra‐structure (Wilson et al. 1987; Bruyas et al. 1995) and metabolic activity (Rieger et al.…”

Section: Freezing Techniquesmentioning

confidence: 99%

“…1991). The alternative CPAs selected had a lower molecular weight that was expected to facilitate their entry into embryonic cells (Bruyas et al. 1995; Bruyas 2011).…”

Section: Freezing Techniquesmentioning

confidence: 99%

“…1995; Bruyas 2011). However, while some CPAs appeared to be relatively non‐toxic, for example, ethylene glycol (Bruyas et al. 1995) and methanol (Bass et al.…”

Section: Freezing Techniquesmentioning

confidence: 99%

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Cryopreservation of Equine Embryos: Current State‐of‐the‐Art

Stout

2012

Reprod Domestic Animals

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Contents During the past 15 years, embryo transfer (ET) has become increasingly widespread within the sport‐horse breeding industry. At present, however, the vast majority (>95%) of horse embryos are transferred fresh or after chilled storage for up to 24 h, whereas cryopreservation is rarely employed despite its obvious potential for simplifying recipient mare management and facilitating long‐term storage and international transport of embryos. A number of inter‐related factors have contributed to the slow development and implementation of equine embryo cryopreservation, and these include the following: (i) the absence of commercially available products for reliably stimulating superovulation; (ii) very poor pregnancy rates following cryopreservation of embryos >300 μm in diameter; (iii) difficulty in recovering embryos at early developmental stages amenable to cryopreservation; and (iv) interembryo variation in susceptibility to cryodamage. However, acceptable success rates (>55% pregnancy) have been reported for both slow‐frozen and vitrified small embryos (<300 μm), and there is renewed interest in cryopreservation, not only in the context of standard ET programmes, but also because it would facilitate pre‐implantation genetic testing and allow wider access to techniques for producing embryos in vitro, such as intracytoplasmic sperm injection and nuclear transfer. This article will review the current status of equine embryo cryopreservation.

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Vitrification of Equine In Vivo-Derived Embryos After Blastocoel Aspiration

Herrera

2020

Cryopreservation and Freeze-Drying Protocols

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Quantitative Analysis of Morphological Modifications of Day 6.5 Horse Embryos after Treatment with Four Cryoprotectants: Differential Effects on Inner Cell Mass and Trophoblast Cells1

Cited by 14 publications

References 46 publications

Cell lineage allocation in equine blastocysts produced in vitro under varying glucose concentrations

Cell lineage allocation in equine blastocysts produced in vitro under varying glucose concentrations

Cryopreservation of Equine Embryos: Current State‐of‐the‐Art

Vitrification of Equine In Vivo-Derived Embryos After Blastocoel Aspiration

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