Microwell bag culture for large-scale production of homogeneous islet-like clusters

Pluripotent stem-cell derived cells can be used for type I diabetes treatment, but we require at least 10⁵–10⁶ islet-like clusters per patient. Although thousands of uniform cell clusters can be produced using a conventional microwell plate, numerous obstacles need to be overcome for its clinical use. In this study, we aimed to develop a novel bag culture method for the production of uniform cell clusters on a large scale (10⁵–10⁶ clusters). We prepared small-scale culture bags (< 10⁵ clusters) with microwells at the bottom and optimized the conditions for producing uniform-sized clusters in the bag using undifferentiated induced pluripotent stem cells (iPSCs). Subsequently, we verified the suitability of the bag culture method using iPSC-derived pancreatic islet cells (iPICs) and successfully demonstrate the production of 6.5 × 10⁵ uniform iPIC clusters using a large-scale bag. In addition, we simplified the pre- and post-process of the culture—a degassing process before cell seeding and a cluster harvesting process. In conclusion, compared with conventional methods, the cluster production method using bags exhibits improved scalability, sterility, and operability for both clinical and research use.

Microwell bag culture for large-scale production of homogeneous islet-like cell clusters. 京都大学プレスリリース. 2022-05-02.