• Developed a two-step monolayer feeder-free protocol for generating iPSC-derived NK cells, offering a more efficient and uniform alternative to embryoid body-based methods.
• iNK cells exhibit key NK cell markers (CD45, CD56, CD16) and functional receptors, with transcriptomic profiles closely resembling peripheral blood NK cells.
• iNK cells demonstrate superior cytotoxicity against cholangiocarcinoma and breast cancer cell lines compared to NK-92 cells in both 2D and 3D tumor spheroid models.
• This approach supports the development of off-the-shelf NK cell products for adoptive immunotherapy, potentially improving clinical scalability and consistency.