3D porous microscaffold promoted hPSC-macrophage large-scale production by facilitating proliferation and self-renewal gene network

Macrophages play crucial roles in various physiological and pathological processes, including immune response and tissue homeostasis. Therefore, the importance of macrophages in preclinical study and clinical application is increasing, necessitating the development of more efficient human pluripotent stem cell(hPSCs) derived macrophage(iMac) differentiation methods to meet the growing demand. Utilizing three-dimensional porous microscaffold (PMS), we have devised a novel approach to induce hPSCs to generate a substantial number of macrophages. This method differs from existing hematopoietic differentiation methods for iMacs. Interestingly, it leads to the formation of a vascular-like hematopoiesis structure resembling the early aorta-gonad-mesonephros(AGM) during embryogenesis. The subsequent event involved the generation of a substantial quantity of macrophages undergoing robust proliferation. Multi-omics data analysis revealed that the 3D microenvironment may generate numerous ligands that promote macrophage proliferation and activate the expression of proliferation and self-renewal genes during iMac generation stage. The subsequent functional experiments demonstrated iMacs can serve as off-the-shelf products for the treatment of infection symptoms caused by drug-resistant Streptococcus pneumoniae infections, and may allow for further innovative cell-based treatment strategies.