3D model to study the immunomodulatory functions of adipocyte/T lymphocytes paracrine crosstalk

Adipose tissue (AT) dysfunction in obesity involves complex interactions among adipocytes, immune cells, endothelial cells and extracellular matrix components. However, most in vitro studies investigating adipocyte-immune interactions rely on simplified two-dimensional systems that incompletely recapitulate human adipose tissue physiology. Here, we developed a patient-derived three-dimensional (3D) human adiposphere model to investigate adipocyte-T-cell crosstalk under controlled metabolic conditions. Adipospheres were generated from the stromal vascular fraction of human subcutaneous AT obtained from donors with clinically distinct metabolic profile. Following 3D adipogenic maturation under normogenic or obesogenic conditions, adipospheres retained donor-specific adipogenic and metabolic characteristics during long-term culture, as demonstrated by differential expression of adipogenic markers, insulin-responsive AKT signaling and lipolytic activity. Compared with normogenic maturation condition, obesogenic maturation induced marked alterations in adiposphere morphology, cytokine secretion and metabolic function. In a second step, mature adipospheres were co-cultured with activated blood-derived T cells using a Transwell-based paracrine system. T-cells co-culture further modulated adiposphere inflammatory and metabolic- related responses. Transcriptomic analyses revealed coordinated regulation of immune- and metabolism related pathways, indicating extensive transcriptional remodeling during adipocyte-T-cell crosstalk. Overall, this modular 3D adiposphere platform reproduces key features of human adipose tissue immunometabolic interactions while preserving donor-specific responses. The system provides a flexible framework for integrating additional cellular components and may facilitate future mechanistic studies and therapeutic screening approaches targeting obesity-associated metabolic and inflammatory disorders. .