Microfluidic Embryo Culture and Analysis

描述（由申请人提供）：本提案是一个设计导向的项目，旨在开发一种新型微流控生物反应器，该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。降低高阶多胎妊娠的发生率，同时保持试管婴儿的整体成功，这是人类试管婴儿的圣杯，生产和鉴定最高质量胚胎的能力将极大地帮助人类试管婴儿。迄今为止，这一目标一直难以实现，至少部分原因是缺乏工具来进行方便可靠的单胚胎操作和分析。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的能力，以及以最小稀释度直接分析胚胎分泌的可溶性生物标志物的培养基的能力。该微流控泵、阀门和传感器的微生物反应器将为胚胎培养提供一个固有的仿生环境，并能在芯片上进行直接的生物标志物分析。微生物反应器将利用计算机控制的集成微流体平台，通过机械微致动器的微通道变形来控制弹性毛细血管内的流体流动。这项探索性资助将监测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具体测试假设，即分析选定的生化标记将能够预测哪8个细胞胚胎将继续产生健康的囊胚。该试验将主要用于设备验证和概念可行性，但也具有临床相关性。目前的趋势是将胚胎培养到囊胚期，并移植两个形态“最好”的囊胚。然而，最近有两篇使用小鼠胚胎的报告独立地证明，将培养延长到囊胚期会导致异常的遗传印记，并改变出生后的发育、生长、生理和行为。病例报告和研究还表明，体外培养到囊胚阶段与MZ双胞胎之间存在普遍关联。该提案将通过开发新的非侵入性方法来选择具有最大植入潜力的胚胎，以最少的操作和培养来解决这些问题。

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.