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首页> 外文期刊>Cell and Tissue Research >Vitrification of goat preantral follicles enclosed in ovarian tissue by using conventional and solid-surface vitrification methods
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Vitrification of goat preantral follicles enclosed in ovarian tissue by using conventional and solid-surface vitrification methods

机译：使用常规和固体表面玻璃化方法玻璃化封闭在卵巢组织中的山羊窦前卵泡

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摘要

Caprine preantral follicles within ovarian fragments were exposed to or vitrified in the presence of sucrose, dimethyl sulfoxide (DMSO), ethylene glycol (EG), or various combinations thereof. The fragments were cryopreserved by using either a conventional (CV) or a solid-surface vitrification (SSV) protocol, and the cryoprotectants were removed by equilibrating vitrified ovarian fragments in “warming solution” consisting of minimum essential medium and heat-inactivated fetal calf serum (MEM+) followed by washes in MEM+ with or without sucrose. Histological analysis of follicle integrity showed that the percentages of normal follicles in ovarian fragments vitrified in sucrose mixed with EG and/or DMSO (CV method) or mixed with EG or DMSO (SSV method) followed by washes in MEM+ plus sucrose were similar to those of controls (ovarian fragments fixed without previous vitrification). Unlike for MEM+ (supplemented or unsupplemented by sucrose) and DMSO followed by washes in the absence of sucrose, the percentages of normal follicles found after exposure to cryoprotectant did not significantly differ from that found after vitrification, indicating that follicular degeneration was attributable to a toxic effect of cryoprotectants and not to the vitrification procedure. The viability of preantral follicles after the CV and SSV procedures was investigated by using calcein-AM and the ethidium-homodimer as “live” and “dead” markers, respectively. In both tested vitrification procedures, the highest percentages of viable follicles were observed when a mixture of sucrose and EG (70.3% for CV and 72.4% for SSV) was used. Preantral follicles were also vitrified (either by CV or SSV) in sucrose and EG and then cultured for 24 h, after which their viability was compared with that of cultured fresh and uncultured vitrified follicles. The viability of these follicles was maintained after SSV, but not after CV. Thus, the viability of caprine preantral follicles can be best preserved after SSV in a mixture of sucrose and EG, followed by washes in medium containing sucrose.
机译：在蔗糖，二甲基亚砜（DMSO），乙二醇（EG）或它们的各种组合存在下，将卵巢片段中的前肛门前卵泡暴露或玻璃化。通过使用常规（CV）或固体表面玻璃化（SSV）方案将片段冷冻保存，并通过在最小限度的培养基和热灭活的胎牛血清组成的“加温溶液”中平衡玻璃化的卵巢片段来去除冷冻保护剂。 （MEM + ），然后在有或没有蔗糖的MEM + 中洗涤。卵泡完整性的组织学分析表明，在蔗糖与EG和/或DMSO（CV方法）混合或与EG或DMSO（SSV方法）混合的蔗糖中玻璃化的卵巢碎片中正常卵泡的百分比，然后在MEM + 加蔗糖中洗涤与对照的相似（卵巢碎片无需事先玻璃化即可固定）。不同于MEM + （被蔗糖补充或未补充）和DMSO，然后在不存在蔗糖的情况下进行洗涤，与冷冻保护剂接触后发现的正常卵泡百分比与玻璃化后发现的卵泡百分比没有显着差异，这表明卵泡变性是归因于防冻剂的毒性作用，而不是玻璃化过程。通过分别使用钙黄绿素-AM和乙锭-同二聚体作为“活”和“死”标志物，研究了CV和SSV手术后的窦前卵泡的生存能力。在两种测试的玻璃化程序中，当使用蔗糖和EG的混合物（CV为70.3％，SSV为72.4％）时，可观察到的卵泡百分比最高。窦前卵泡也用蔗糖和EG进行玻璃化（通过CV或SSV），然后培养24小时，然后将其活力与培养的新鲜和未培养的玻璃化卵泡进行比较。这些卵泡的活力在SSV后得以维持，而在CV后则没有。因此，SSV后在蔗糖和EG的混合物中可以最好地保留山羊腹腔前卵泡的活力，然后在含蔗糖的培养基中洗涤。

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  • 来源
    《Cell and Tissue Research》 |2007年第1期|167-176|共10页
  • 作者

    R. R. Santos; T. Tharasanit; T. Van Haeften; J. R. Figueiredo; J. R. V. Silva; R. Van den Hurk;

  • 作者单位

    Departmant of Farm Animal Health Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands;

    Department of Equine Sciences Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands;

    Department of Biochemistry and Cell Biology Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands;

    Laboratory of Manipulation of Oocytes Enclosed in Preantral Follicles - LAMOFOPA Faculty of Veterinary Medicine Ceará University Fortaleza CE Brazil;

    Laboratory of Manipulation of Oocytes Enclosed in Preantral Follicles - LAMOFOPA Faculty of Veterinary Medicine Ceará University Fortaleza CE Brazil;

    Departmant of Farm Animal Health Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Preantral follicles; Vitrification; Viability markers; In vitro culture; Caprine;

    机译：窦前卵泡;玻璃化;活力标记;体外培养;山羊;

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