Jones E A V, Crotty D, Kulesa P M, Waters C W, Baron M H, Fraser S E, Dickinson M E
Department of Chemical Engineering, California Institute of Technology, Pasadena, California.
Genesis. 2002 Dec;34(4):228-35. doi: 10.1002/gene.10162.
Due to the internal nature of mammalian development, much of the research performed is of a static nature and depends on interpolation between stages of development. This approach cannot explore the dynamic interactions that are essential for normal development. While roller culture overcomes the problem of inaccessibility of the embryo, the constant motion of the medium and embryos makes it impossible to observe and record development. We have developed a static mammalian culture system for imaging development of the mouse embryo. Using this technique, it is possible to sustain normal development for periods of 18-24 h. The success of the culture was evaluated based on the rate of embryo turning, heart rate, somite addition, and several gross morphological features. When this technique is combined with fluorescent markers, it is possible to follow the development of specific tissues or the movement of cells. To highlight some of the strengths of this approach, we present time-lapse movies of embryonic turning, somite addition, closure of the neural tube, and fluorescent imaging of blood circulation in the yolk sac and embryo.
由于哺乳动物发育的内在特性，所进行的许多研究具有静态性质，并且依赖于发育阶段之间的插值。这种方法无法探索正常发育所必需的动态相互作用。虽然旋转培养克服了胚胎难以接近的问题，但培养基和胚胎的持续运动使得观察和记录发育变得不可能。我们开发了一种用于对小鼠胚胎发育进行成像的静态哺乳动物培养系统。使用该技术，可以维持18 - 24小时的正常发育。基于胚胎转动速率、心率、体节添加以及几个总体形态特征对培养的成功进行了评估。当该技术与荧光标记相结合时，就有可能追踪特定组织的发育或细胞的运动。为了突出这种方法的一些优势，我们展示了胚胎转动、体节添加、神经管闭合以及卵黄囊和胚胎中血液循环的荧光成像的延时电影。
