Microfluidic Embryo Culture and Analysis

描述（由申请人提供）：该提案是一个设计导向项目，旨在开发一种新型微流体生物反应器，该生物反应器将在模拟生理条件下培养多个单胚胎，同时对胚胎质量的生化标志物进行实时监测。在保持整体试管婴儿成功率的同时减少高阶多胎妊娠的发生率，Irate是人类试管婴儿的圣杯，并且能够生产和识别最高质量的胚胎将大大有助于。迄今为止，这一目标一直难以实现，至少部分原因是缺乏进行方便可靠的单胚胎操作和分析的仪器。由于单个胚胎产生的生物标志物数量很少，因此可靠的定量取决于在非常小体积的液体中培养胚胎并直接分析培养基中由胚胎分泌的可溶性生物标志物的能力。所提出的具有微流体泵、阀和传感器的微生物反应器将为胚胎培养提供固有的仿生环境，并使芯片上的直接生物标志物分析成为可能。微生物反应器将利用计算机控制的集成微流体平台，该平台通过机械微致动器使微通道变形来控制弹性体毛细管内的流体流动。在这项探索性资助中将监测的小鼠胚胎健康的生物标志物是胚胎代谢物、自分泌因子和胚胎健康的胚胎表面生物标志物。胚胎微生物反应器将用于专门测试以下假设：选择的生化标志物的分析将能够预测哪个8细胞胚胎将继续产生健康的囊胚。本试验主要用于器械确认和概念可行性，但也具有临床相关性。目前的趋势是将胚胎培养到囊胚阶段，并将两个形态学上“最好”的囊胚移植。然而，最近，两个使用小鼠胚胎的报告已经独立地证明，延长培养到囊胚阶段会导致异常的遗传印记和出生后发育，生长，生理和行为的改变。病例报告和研究还表明，体外培养到囊胚阶段与MZ双胎之间存在一般关联。该提案将通过开发新的非侵入性方法来解决这些问题，这些方法可以选择具有最大植入潜力的胚胎，同时操作和培养量最少。

DESCRIPTION (provided by applicant): This proposal is a design-directed project to develop a novel microfluidic bioreactor that will culture multiple single embryos under simulated physiological conditions while simultaneously performing real-time monitoring of biochemical markers of embryo quality. Reducing the incidence of high-order multiple pregnancies while maintaining the overall IVF success Irate is a holy grail of human IVF and would be greatly assisted by the ability to produce and identify the highest quality embryos. This goal has been elusive to date due, at least in part, to the lack of instrumentation to perform convenient and reliable single embryo manipulation and analysis. Because of the small quantities of biomarkers produced by single embryos, reliable quantification hinges on the ability to culture the embryos in very small volumes of fluid and to directly analyze the culture media for soluble biomarkers secreted by embryos with minimal dilution. The proposed microbioreactor with microfluidic pumps, valves, and sensors will provide an inherently biomimetic milieu for embryo culture as well as enable direct biomarker analysis on chip. The microbioreactor will utilize a computer-controlled integrated microfluidics platform that controls fluid flow inside elastomeric capillaries by deformation of the microchannels with mechanical microactuators. The biomarkers of mouse embryo health that will be monitored in this exploratory grant are embryo metabolites, autocrine factors, and embryo surface biomarker of embryo health. The embryo microbioreactor will be used to specifically test the hypothesis that analysis of select biochemical markers will enable prediction of which 8 cell embryo will proceed to produce healthy blastocysts. This test will mainly serve the purpose of device validation and concept feasibility but it also has clinical relevance. A current trend is to grow embryos to the blastocyst stage and transfer the two morphologically "best" blastocyst. Recently, however, two reports using mouse embryos have independently demonstrated that extended culture to the blastocyst stage causes aberrant genetic imprinting and altered postnatal development, growth, physiology and behavior. Case reports and studies also suggest a general association between in vitro culture to the blastocyst stage and MZ twinning. This proposal will address these issues by developing novel non-invasive means of selecting embryos with the greatest implantation potential, with the least amount of manipulation and culture.