Microfluidic Embryo Culture and Analysis

描述(申请人提供)：该方案是一个设计指导的项目，旨在开发一种新型的微流控生物反应器，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。在保持总体试管受精成功率的同时，减少高位多胎妊娠的发生率是人类试管受精的一个圣杯，生产和识别最高质量胚胎的能力将极大地帮助这一目标。到目前为止，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记物数量很少，可靠的量化取决于能否在非常少量的液体中培养胚胎，并以最小的稀释度直接分析胚胎分泌的可溶性生物标记物的培养液。所提出的具有微流体泵、阀门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，并能够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，通过机械微执行器使微通道变形来控制弹性毛细血管内的流体流动。在这项探索性拨款中将监测的小鼠胚胎健康的生物标记物是胚胎代谢物、自分泌因子和胚胎健康的胚胎表面生物标记物。胚胎微生物反应器将被用来专门测试这样的假设，即对选定的生化标记的分析将能够预测哪8个细胞胚胎将继续产生健康的囊胚。这项测试将主要服务于设备验证和概念可行性的目的，但也具有临床意义。目前的一种趋势是将胚胎培养到囊胚期，然后将两个形态上最好的囊胚移植到囊胚期。然而，最近，两份使用小鼠胚胎的报告独立地证明，延长培养到囊胚阶段会导致异常的基因印记，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，体外培养到囊胚期和MZ双胞胎之间存在普遍的联系。这项建议将通过开发新的非侵入性方法来解决这些问题，以最少的操作和培养选择具有最大植入潜力的胚胎。

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.