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

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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.