Direct Differentiation of Human Adult Adipose Tissue into Multilineage Functional Organoids

Current strategies for generating organoids typically relay on intricate in vitro manipulation of dissociated stem cells including the isolation, expansion, and genetic processing of cells, which pose safety and scalability challenges for therapeutic translation. The direct generation of organoids from readily accessible tissues may offer a solution to these issues. Herein, we demonstrate the direct generation of functional organoids representing all three germ layers from human adult adipose tissue without any single-cell manipulation steps. Specifically, we generated reaggregated microfat (RMF) tissues from human adipose tissue using a specialized suspension culture system. These RMF tissues can be converted into mesodermal bone marrow organoids capable of reconstituting human normal hematopoiesis in immunodeficient mice; endodermal insulin-producing organoids able to reverse hyperglycemia in STZ-induced diabetic mice; and ectodermal nervous-like tissues resembling neurons and neuroglial cells. These findings highlight the potential of human adult adipose tissue as a safe and scalable platform for generating diverse clinically-relevant organoids for regenerative therapy.