The generation of brown and white adipose organoids from induced pluripotent stem cells through differentiation into...

Adipose tissue is a highly heterogeneous endocrine organ with essential roles in energy storage, metabolic regulation, and thermogenesis. Here, brown adipose (BA) and white adipose (WA) organoids were generated from induced pluripotent stem cells (iPSCs) via differentiation into a neuromesodermal progenitor (NMP) state for the first time, and brown and white differentiation was confirmed by uncoupling protein 1 (UCP1) and fatty acid binding protein 4 (FABP4) expression. Morphological, histological, and ultrastructural analyses revealed depot-specific lipid droplet morphologies. Fatty-acid profiling revealed depot-like lipid signatures, and elevated triglycerides across lipid classes confirmed robust lipid accumulation in the organoids. Notably, small-molecule treatment used for obesity screening induced tissue-relevant responses in adipose organoids, highlighting their potential for drug testing. Moreover, liquid chromatography-tandem mass spectrometry (LC-MS/MS) proteomics analysis revealed distinct programs associated with brown and white adipose-like states, including mitochondrial/thermogenic signatures in BA organoids and enrichment of endomembrane-trafficking pathways in WA organoids. Furthermore, transplantation into CD1 nude mice demonstrates tissue integration with enhanced vascularization of the graft. iPSC-derived adipose organoids reproduce key depot-like phenotypes, lipid composition, proteomic programs, and treatment responsiveness, providing a scalable human platform for metabolic research and future patient-oriented screening.