Standardized generation of human iPSC-derived hematopoietic organoids and macrophages utilizing a benchtop bioreactor platform under fully defined conditions

There is a significant demand for intermediate-scale bioreactors in academic and industrial institutions to produce cells for various applications in drug screening and/or cell therapy. However, the application of these bioreactors in cultivating hiPSC-derived immune cells and other blood cells is noticeably lacking. To address this gap, we have developed a xeno-free and chemically defined intermediate-scale bioreactor platform, which allows for the generation of standardized human iPSC-derived hematopoietic organoids and subsequent continuous production of macrophages (iPSC-Mac). Human iPSC-Mac derived from three different iPSC lines were harvested as described in the study. Viable cells were purified by fluorescence activated cell sorting (FACS) on basis of scatter properties and exclusion of 4′,6-diamidino-2-phenylindole (DAPI) positive events, which has been added immediately before sorting (0.1 µg/ml final concentration).