FMSG: Bio: Advancing Extracellular Vesicle Biomanufacturing of CRISPR-Edited Human iPSC-derived MSCs with Next-Generation Purification

间充质干细胞（MSCs）产生的细胞外囊泡（EVs）因其多方面的治疗潜力而备受关注。然而，EV治疗的实际应用面临的一个巨大挑战是治疗性干细胞的EV产量低。为了开发先进的电动汽车生物制造工艺以满足临床需求，该项目旨在通过整合人类诱导多能干细胞（hiPSCs）来扩大供体细胞来源的可扩展性，基因组工程来扩大电动汽车生物发生的可扩展性，以及先进的纳米膜技术来扩大电动汽车纯化的可扩展性，从而实现msc衍生电动汽车的规模化生产。此外，该项目旨在整合研究、教育、工业和多样性，重点是在多个层面上加强学生的研究机会。它将通过在两个校区开发新的合作课程，为研究生和本科生提供研究机会，并利用两个学院的现有项目开展外展活动来实现。紧密的产业合作也将为研究生提供行业实习机会。该项目不仅解决了治疗性电动汽车生物制造中供体干细胞来源的可扩展性问题，而且为电动汽车生物学领域提供了新的见解，并为电动汽车纯化的瓶颈提供了新的解决方案。在此，本研究汇集了一个跨学科的研究团队，以(1)产生稳定的hiPSC系，合成EV生物发生和生物活性的助推器，(2)建立一个无血清分化的围产期组织样iMSCs作为一个强大的EV工厂，(3)优化化学启动条件，增强细胞代谢活性，促进EV生产，(4)引入纳米袋膜技术，可扩展EV纯化，提高产量和产量。与其他标准技术相比。hiPSC技术、基因组工程和膜纳米制造的整合为生产具有高可扩展性、理想功能和临床相关性的治疗性电动汽车提供了巨大的潜力，为未来电动汽车生物制造的进步提供了巨大的潜力。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Extracellular vesicles (EVs) produced by mesenchymal stem cells (MSCs) have been highlighted for their multifaceted therapeutic potentials. However, a great challenge of practical applications of EV therapy is the low production of EVs from therapeutic stem cells. To develop an advanced EV biomanufacturing process to meet the clinical need, this project aims to enable scale-up production of MSC-derived EVs by integrating human induced pluripotent stem cells (hiPSCs) for scalability in donor cell source, genome engineering for scalability in EV biogenesis, and advanced nano-membrane technology for scalability in EV purification. Furthermore, this project is designed to integrate research, education, industry, and diversity with an emphasis on strengthening research exposure to the students at multiple levels. It will be achieved by developing new collaborative classes across two campuses, providing research opportunities to graduate and undergraduate students, and leveraging existing programs at both institutes for outreach activities. The close industrial collaboration will also present industry internship opportunities to the graduate students.This project not only addresses the scalable issues in donor stem cell sources for therapeutic EV biomanufacturing, but also provides new insights to the field of EV biology and new solutions to the bottleneck of EV purification. Herein, this research brings together an interdisciplinary research team to (1) generate a stable hiPSC line with synthetic boosters of EV biogenesis and bioactivity, (2) establish a serum-free differentiation of perinatal tissue-like iMSCs as a potent EV factory, (3) optimize chemical priming conditions to enhance cell metabolic activities for boosting the EV production, and (4) introduce the nanopocket membrane technique for scalable EV purification with improved throughput and yield, comparing to the other standard techniques. Integration of hiPSC technology, genome engineering and membrane nanofabrication offers great potential to produce therapeutic EVs with high scalability, desired functions, and clinical relevance for future advancement of EV biomanufacturing.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.