Winn Louise M, Tung Emily W Y
Department of Pharmacology and School of Environmental Studies, Queen's University, Kingston, Ontario, Canada.
Methods Mol Biol. 2009;550:241-9. doi: 10.1007/978-1-60327-009-0_15.
The mouse embryo culture technique is a valuable tool for assessing embryotoxicity of exogenous compounds as it excludes any confounding maternal and placental effects, allows for the selection of embryos that are at similar stages of development, and permits the control of exposure concentrations of exogenous agents and modifiers of interest. This chapter will use the anticonvulsant drug valproic acid as a model teratogen to describe the mouse embryo culture in detail. Briefly, mice are bred and the presence of a vaginal plug in a female mouse indicates gestational day (GD) 1. On GD 9 embryos are explanted from pregnant dams and embryos that are at similar stages of development (4-6 somite pairs) are cultured in CO(2) saturated male rat serum for 24 h at 37 degrees C. After 24 h embryonic morphological and developmental parameters, including anterior neuropore closure, are evaluated using a dissecting microscope. Additional biochemical analysis, including molecular approaches to assess embryonic signal transduction, as well as some limitations of the technique will also be discussed.
小鼠胚胎培养技术是评估外源性化合物胚胎毒性的一项重要工具，因为它排除了母体和胎盘的任何混杂效应，能够选择处于相似发育阶段的胚胎，并可控制外源性物质及相关调节剂的暴露浓度。本章将以抗惊厥药物丙戊酸作为典型致畸剂，详细描述小鼠胚胎培养过程。简要来说，将小鼠进行交配，雌性小鼠出现阴栓表明妊娠第1天（GD1）。在妊娠第9天，从怀孕母鼠体内取出胚胎，将处于相似发育阶段（4 - 6对体节）的胚胎在37℃、含二氧化碳饱和的雄性大鼠血清中培养24小时。24小时后，使用解剖显微镜评估胚胎的形态和发育参数，包括前神经孔闭合情况。还将讨论其他生化分析，包括评估胚胎信号转导的分子方法，以及该技术的一些局限性。
