Preantral follicle culture as a novel in vitro assay in reproductive toxicology testing in mammalian oocytes
Preantral follicle culture as a novel in vitro assay in reproductive toxicology testing in mammalian oocytes
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DOI：
10.1093/mutage/geg040
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发表时间：
2004-01-01
期刊：
影响因子：
2.7
通讯作者：
Eichenlaub-Ritter, U
中科院分区：
文献类型：
--
作者：
Sun, FY;Betzendahl, I;Eichenlaub-Ritter, U
人类最常见的遗传病，三体，主要是由卵子发生过程中染色体分离的错误引起的。分离的小鼠卵母细胞在体外恢复减数分裂并进展到中期II期，最近已被用于评估卵母细胞的靶标、非优生潜能和对化学暴露的敏感性。为了将体外成熟试验扩展到卵子发生的早期阶段，建立了小鼠腔前卵泡培养的体外试验方法。它可以识别环境化学物质对体细胞、卵泡和膜细胞的直接和间接影响，这些影响可能导致卵母细胞的生长、成熟和染色体分离的紊乱。在严格控制的条件下，将青春期前雌性小鼠的早期腔前卵泡在微滴中培养12天。在重组人促性腺激素和表皮生长因子的生理性排卵刺激下，卵泡包裹的卵母细胞在16h内恢复成熟，发育到中期II，并在体外排卵。这些在体外生长和成熟的卵母细胞具有正常的桶形纺锤体和排列良好的染色体。它们的染色体在后期I高保真地分离。模型去雄秋水仙素以剂量依赖的方式诱导这些体外培养的卵泡封闭的卵母细胞减数分裂停滞和非整倍体，与体内试验相当。因此，腔前卵泡培养提供了一种有效和可靠的方法来评估环境诱变剂、药物和潜在的内分泌干扰物对雌性减数分裂保真度的影响。
The most common genetic disorder in humans, trisomy, is caused predominantly by errors in chromosome segregation during oogenesis. Isolated mouse oocytes resuming meiosis and progressing to metaphase II in vitro have recently been used to assess targets, aneugenic potential and sensitivity of oocytes to chemical exposures. In order to extend in vitro maturation tests to earlier stages of oogenesis, an in vitro assay with mouse preantral follicle cultures has been established. It permits the identification of direct and also indirect effects of environmental chemicals on the somatic compartment, the follicle and theca cells, that may lead to disturbances of oocyte growth, maturation and chromosome segregation. Early preantral follicles from prepubertal female mice are cultured in microdroplets for 12 days under strictly controlled conditions. The follicle-enclosed oocytes resume maturation, develop to metaphase II and become in vitro ovulated within 16 h after a physiological ovulatory stimulus with recombinant human gonadotrophins and epidermal growth factor. These oocytes grown and matured in vitro possess normal barrel-shaped spindles with well-aligned chromosomes. Their chromosomes segregate with high fidelity during anaphase I. The model aneugen colchicine induced a meiotic arrest and aneuploidy in these in vitro grown, follicle-enclosed oocytes in a dose-dependent manner, comparable to in vivo tests. Therefore, preantral follicle culture appears to provide an effective and reliable method to assess the influences of environmental mutagens, pharmaceutical agents and potentially endocrine disrupting chemicals on the fidelity of female meiosis.