DEVELOPMENT OF A HUMAN PLACENTA VILLOUS MICROPERFUSION

对人体胎盘的体内接触限制和 人类胎儿的血液循环导致了一些 实验模型，虽然在某些方面非常有用， 研究，不能充分代表体内胎盘功能。这些 模型可以分为两种类型。第一种是那些，比如小学。 培养的滋养层，这使得精确的实验规格 条件和允许详细观察，但缺乏结构关系 和体内组织的表型表达。第二种是那些 保持结构拓扑结构，并反映环境的影响， 子宫，如胎盘在体外小叶灌注，但不允许 在细胞水平上的操作或观察。随着更多信息 关于胎盘的分子和细胞功能， 越来越重要的是找到一种方法， 可以在组织模型中检查经胎盘的离子或水转运， 确定它们如何在这个更复杂的多组分系统中运行。是 因此，有必要整合这两种模式，并开发一种新的方法， 其中可以在原代组织中观察到详细的细胞功能。目标 该项目的一个重要目的是开发一种新的模型， 微灌注，这将使这些调查。在这个模型中， 绒毛膜绒毛树的片段将被插管并灌注通过 胎儿循环，同时灌注碎片， 模拟母体循环。这将在以下阶段进行 一个落射荧光显微镜，使观察荧光探针在 合胞体层或胎儿脉管系统中，通过设计方法来保持， 管状和灌注绒毛碎片。第二个目标是验证 绒毛灌注通过测量生化稳定性和结构 完整该模型将为新的研究提供基础， 胎盘运输、代谢、信号传导和结构生物学。

The restrictions on in vivo access to the human placenta and the human fetal circulation have resulted in the development of a number of experimental models, which, while extremely useful for some aspects of investigation, fail to represent placental function in vivo adequately. These models can be divided into two types. The first are those, such as the primary cultured trophoblast, which enable specification of precise experimental conditions and allow detailed observation but lack the structural relationships and phenotypic expression of the tissue in vivo. The second are those which retain the structural topology and reflect the effects of the environment in utero, such as the placental in vitro lobule perfusion, but which do not permit manipulation or observation at the cellular level. As more information is obtained concerning molecular and cellular function of the placenta, it is becoming increasingly important to find a way in which processes such as transplacental ion or water transport can be examined in a tissue model to determine how they operate in this more complex, multi-component system. It is necessary therefore to integrate the two model types and develop a new approach in which detailed cellular function can be observed in primary tissue. The goal of this project is to develop a new model, the chorionic villous microperfusion, which will enable these investigations. In this model a small fragment of the chorionic villous tree will be cannulated and perfused through the fetal circulation, while the fragment is simultaneously superfused, simulating the maternal circulation. This will be carried out on the stage of an epifluoresence microscope, enabling observation of fluorescent probes in the syncytial layer or in the fetal vasculature, by devising methods to hold, cannulate and perfuse villous fragments. The second aim is to validate the villous perfusion through measurements of biochemical stability and structural integrity. This model will provide the basis for new investigations in placental transport, metabolism, and signaling and structural biology.