Division of Veterinary Anatomy and Histology, Department of Animal Sciences, Faculty of Veterinary Medicine, University of Milan, Milan, Italy.
Fertil Steril. 2010 Jan;93(1):229-38. doi: 10.1016/j.fertnstert.2008.09.078. Epub 2008 Nov 1.
To evaluate the use of fresh or frozen bovine oocytes as an animal model for reconstructing artificial gametes by germinal vesicle transplantation (GVT), to study nucleocytoplasmic interaction and define clinical procedures for ooplasm donation in humans.
Prospective experimental study.
University-based experimental laboratory.
ANIMAL(S): Bovine oocytes from slaughterhouse ovaries.
INTERVENTION(S): A total of 446 gametes were reconstructed from fresh immature oocytes; nuclear and cytoplasmic competencies were analyzed through the assessment of meiotic progression and cytoskeleton reorganization; embryonic developmental capability was evaluated after parthenogenetic activation of metaphase II (MII) reconstructed oocytes. Furthermore, the distribution of mitochondria in karyoplast and cytoplast in grafted oocytes was studied. Finally, meiotic and developmental competencies were determined in 199 gametes reconstructed from vitrified immature oocytes.
MAIN OUTCOME MEASURE(S): Maturational and developmental rate of reconstructed oocytes, cytoskeleton organization, and mitochondrial distribution.
RESULT(S): Gametes reconstructed from either fresh or cryopreserved immature oocytes showed similar meiotic competence (41.6% vs. 37.7%, respectively). All reconstituted oocytes that reached MII displayed a normal distribution of cytoskeletal elements. Embryonic developmental capability was higher in oocytes derived from fresh than from cryopreserved gametes (30.8% vs. 8.1%, respectively). Finally, oocyte centrifugation was effective in obtaining karyoplasts with <5% of mitochondria.
CONCLUSION(S): Cows can provide a suitable organism model to develop GVT technique in both research and clinical settings as well as in fertility preservation programs.
评估使用新鲜或冷冻牛卵母细胞作为通过卵母细胞胞质内单精子注射（GVT）重建人工配子的动物模型，研究核质相互作用，并确定人类卵质捐赠的临床程序。
前瞻性实验研究。
以大学为基础的实验实验室。
来自屠宰场卵巢的牛卵母细胞。
共从新鲜未成熟卵母细胞中重建了 446 个配子；通过评估减数分裂进程和细胞骨架重组来分析核和细胞质的能力；通过对中期 II（MII）重构卵母细胞进行孤雌激活来评估胚胎发育能力。此外，研究了移植卵母细胞中核质体和细胞质体中线粒体的分布。最后，确定了从玻璃化未成熟卵母细胞中重建的 199 个配子的减数分裂和发育能力。
重构卵母细胞的成熟和发育率、细胞骨架组织和线粒体分布。
从新鲜或冷冻未成熟卵母细胞中重建的配子显示出相似的减数分裂能力（分别为 41.6%和 37.7%）。所有达到 MII 的重构卵母细胞均显示出正常的细胞骨架元素分布。来自新鲜卵母细胞的胚胎发育能力高于来自冷冻卵母细胞的胚胎发育能力（分别为 30.8%和 8.1%）。最后，卵母细胞离心可以有效地获得线粒体含量<5%的核质体。
牛可以为在研究和临床环境中以及在生育力保存计划中开发 GVT 技术提供合适的动物模型。
