OvaSafe: A microfluidic device enabling automated oocyte vitrification

OvaSafe：一种微流体装置，可实现卵母细胞自动玻璃化冷冻

• 批准号: 10892324
• 负责人: Ismail Emre Ozkumur
• 金额: $ 107万
• 依托单位: AUTOIVF, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10892324
• 关键词: OvaSafe; microfluidic; device; enabling; automated

Abstract In the United States, the total number of In-vitro Fertilization (IVF) clinics has stayed relatively stagnant through the past decade, and the IVF-cycles per million women remains significantly lower than in other developed countries. The greatest challenge for widespread use of IVF includes its high cost, driven by the need for complex and expensively equipped IVF laboratories and highly-trained embryologists. The high cost and accessibility of IVF clinics is the leading reason why couples are unable to undergo or delay IVF treatment and/or egg banking. Our company’s goal is to automate various functions within an embryology lab to decrease the cost and improve the overall accessibility of IVF treatments and fertility preservation. We are aiming to achieve this by reducing the equipment and personnel requirements so as to pave the way towards de-centralization of IVF. Towards this goal, our proposed new technology “OvaSafe” will facilitate vitrification of oocytes prior to them being transferred to central embryology labs for fertilization or long-term storage facilities for preservation. OvaSafe will complement the ongoing project, OvaReady, which automatically prepares oocytes for vitrification. OvaSafe will utilize microfluidic devices to load cryoprotectant agents (CPA) rapidly and gradually to oocytes, and automatically transfer the oocytes into silica capillary carriers prior to them being plunged into liquid nitrogen to complete the vitrification process. In addition to improving the accessibility by enabling processing of gametes at satellite clinics, performing gradual CPA loading using microfluidic devices will reduce the CPA exposure, toxicity, and osmotic stress to oocytes. Automated CPA loading and utilization of a closed vitrification carrier will eliminate variations between operators and clinics. Although the scope of this proposal is limited to oocyte vitrification, the technology can also be applicable to embryo vitrification in the future. In our preliminary studies, we demonstrated that gradual CPA loading using microfluidics and silica capillary vitrification are both practical methods which can improve vitrification outcomes. We showed that gradual CPA loading decreases the total CPA exposure to oocytes, and reduces the osmotic stress driven by cell shrinkage which is a response to standard step-wise CPA loading protocols. We also demonstrated that silica capillaries can achieve ultra-rapid cooling and warming rates that are comparable to standard open carrier methods, and the vitrification outcomes with silica capillaries are comparable or superior to standard methods. In the proposed work, we will build on these innovations and integrate both methods in a simple, reliable, and closed system to automate the oocyte vitrification process. Through this Fast-Track application, we propose and plan to complete the following aims in this order: develop a microfluidic CPA loading device; modify CPA loading device for automated oocyte loading to silica capillaries; have plastic microfluidic devices and customized silica capillaries manufactured to improve usability; design and build an automated control system; demonstrate success of the automated workflow with vitrification of mouse and donated human oocytes.

抽象的 在美国，维特罗施肥（IVF）诊所的总数已经相对停滞 过去十年，每百万妇女的IVF周期均显着低于其他发达的发达 国家。 IVF宽度使用的最大挑战包括其高成本，这是由于需要复杂的需求而驱动 以及全面等效的IVF实验室和训练有素的胚胎学家。高成本和可访问性 IVF诊所的主要原因是夫妻无法接受或延迟IVF治疗和/或鸡蛋的主要原因 银行业。我们公司的目标是在胚胎学实验室中自动化各种功能以降低成本 并改善IVF治疗和生育能力的总体可及性。我们的目标是实现这一目标 通过减少设备和人员要求，以铺平IVF偏心化的道路。 为了实现这一目标，我们提出的新技术“ ovasafe”将促进卵母细胞的剥离 被转移到中央胚胎学实验室进行施肥或长期存储设施以保存。 Ovasafe将完成正在进行的项目Ovaready，该项目会自动准备卵母细胞以进行玻璃化。 Ovasafe将利用微流体设备快速加载冷冻保护剂（CPA） 卵母细胞，然后自动将卵母细胞转移到二氧化硅毛细管中 进入液氮以完成玻璃化过程。除了通过启用提高可访问性 在卫星诊所处理游戏的处理，使用微流体设备进行CPA级负载将减少 CPA暴露，毒性和渗透应激对卵母细胞。自动化的CPA加载和封闭的利用 玻璃化载体将消除操作员和诊所之间的变化。尽管该提议的范围是 该技术仅限于卵母细胞玻璃化，将来也适用于胚胎玻璃化。 在我们的初步研究中，我们证明了使用微流体和二氧化硅毛细管的CPA载荷 玻璃化都是可以改善玻璃化结果的实际方法。我们证明了CPA级 加载可减少CPA暴露于卵母细胞，并减少由细胞收缩驱动的渗透压 这是对标准阶梯CPA加载协议的响应。我们还证明了二氧化硅毛细血管 可以实现与标准开放式载体方法相媲美的超优化冷却速率，并且 二氧化硅毛细血管的玻璃化结果可与标准方法相当或优越。在提议中 工作，我们将以这些创新为基础，并将两种方法集成到一个简单，可靠且封闭的系统中 自动化卵母细胞玻璃化过程。通过此快速轨道应用程序，我们建议并计划完成 以下目的是：开发微流体CPA加载设备；修改CPA加载设备 自动卵母细胞加载到二氧化硅毛细血管上；具有塑料微流体设备和定制的二氧化硅毛细管 制造以提高可用性；设计和构建自动控制系统；展示成功的成功 自动化的工作流量，玻璃体的玻璃化并捐赠了人类卵母细胞。