Identifying and optimizing critical process parameters for large-scale manufacturing of iPSC derived insulin-producing β-cells

作者：Haneen Yehya, Alexandra Wells, Michael Majcher, Dhruv Nakhwa, Ryan King, Faruk Kerem Şentürk, Roshan Padmanabhan, Jan Jensen, Michael A. Bukys · 发表于：Stem Cell Research & Therapy · 年份：2024 · DOI：10.1186/s13287-024-03973-0 · 被引用次数：6 · 研究领域：Pancreatic function and diabetes、Pluripotent Stem Cells Research、Diabetes Management and Research

BACKGROUND: Type 1 diabetes, an autoimmune disorder leading to the destruction of pancreatic β-cells, requires lifelong insulin therapy. Islet transplantation offers a promising solution but faces challenges such as limited availability and the need for immunosuppression. Induced pluripotent stem cells (iPSCs) provide a potential alternative source of functional β-cells and have the capability for large-scale production. However, current differentiation protocols, predominantly conducted in hybrid or 2D settings, lack scalability and optimal conditions for suspension culture. METHODS: We examined a range of bioreactor scaleup process parameters and quality target product profiles that might affect the differentiation process. This investigation was conducted using an optimized High Dimensional Design of Experiments (HD-DoE) protocol designed for scalability and implemented in 0.5L (PBS-0.5 Mini) vertical wheel bioreactors. RESULTS: A three stage suspension manufacturing process is developed, transitioning from adherent to suspension culture, with TB2 media supporting iPSC growth during scaling. Stage-wise optimization approaches and extended differentiation times are used to enhance marker expression and maturation of iPSC-derived islet-like clusters. Continuous bioreactor runs were used to study nutrient and growth limitations and impact on differentiation. The continuous bioreactors were compared to a Control media change bioreactor showing metabolic shifts and a more β-cel...