Porous microscaffold enables spontaneous hematopoietic cell differentiation and continuous macrophage production from hPSCs for therapy.

Abstract

Macrophages play critical roles in various physiological and pathological processes, yet their scalable production for therapeutic applications remains a challenge. Here, we present a porous microscaffold (PMS) that enables robust hematopoietic differentiation from human pluripotent stem cell (hPSC)-derived mesoderm. Within PMS, mesoderm cells self-organize into vascular structures reminiscent of the early aorta-gonad-mesonephros (AGM) niches, promoting efficient endothelial-to-hematopoietic transition with minimal cytokine supplementation. Additionally, endothelial-cell-modified PMS enriches paracrine signals and activates key genes involved in macrophage proliferation and self-renewal. Multi-omics analyses delineate the gene network driving enhanced macrophage expansion within this 3D microenvironment. Functionally, PMS-derived macrophages exhibit potent therapeutic potential by effectively alleviating drug-resistant Streptococcus pneumoniae infections in mice. These results establish PMS as a cost-effective and scalable platform for producing functional macrophages, paving the way for off-the-shelf cell-based therapies.