Towards a scalable, closed and automated platform for the production of cost-efficient allogeneic cell therapies: showcase of an exemplar iNK process

Induced pluripotent stem cell (iPSC)-derived therapies offer promising "off-the-shelf" allogeneic treatments for numerous chronic conditions. However, their efficacy requires high doses exceeding ≥10⁹ cells, which are not suited to traditional 2D processes that are typically open and poorly scalable technologies. Successful commercialization hinges on developing efficient processing platforms for large-scale manufacture. Here, we introduce a closed, scalable, semi-automated upstream process for iPSC expansion and differentiation into Natural Killer (iNK) cells using stirred tank reactors (STR). Unlike traditional methods relying on periodic settling for medium exchanges, our method employs an acoustic filtration system to continuously exchange medium and control aggregate size, crucial for expansion and differentiation while preserving pluripotency. Our results demonstrate reproducible high-density cultures (>10⁶ cells/mL) with controlled aggregate diameter (100-200 µM).

In collaboration with Plasticell, we have scaled up a process for manufacturing iPSC-derived NK cells. We successfully integrated our iPSC expansion platform and enhanced Plasticell's NK differentiation process using a scalable STR system. This advancement yields a pure population of CD45+/CD56+ NK cells (>80%) with increased yield per starting iPSC compared to manual methods. These cells exhibit mature NK cell markers and demonstrate cytotoxicity against lung and breast cancer cell lines in vitro.

We believe our work represents the first end-to-end process for generating iNK cells in an STR system. This integrated approach accelerates patient access to transformative allogeneic therapies, marking a significant contribution to the field.