From sebocytes to skin engineering human sebaceous organoids within a reconstructed dermal matrix

Introduction: systems fail to capture these features simultaneously. This work reports the generation of a functional three-dimensional (3D) sebaceous gland organoid embedded within a matrix and designed to reproduce the structural and functional properties of native sebaceous tissue. Methods: Using SZ95 sebocytes, we first established baseline responses in monolayer conditions using linoleic acid (LA) stimulation and pharmacological inhibition with retinol (RTN) and capsaicin (CPS). We then reconstructed a 3D organoid model in which a sebocyte-rich lipidogenic compartment was spatially organized within a matrix-supported microenvironment and associated with an outer stromal-like component. The model was evaluated using morphological, ultrastructural, and immunofluorescence analyses. To further define the mode of action of sebogenic activation in 3D, we analyzed genes related to extracellular matrix remodeling and tissue adaptation. Results: In both monolayer and 3D models, LA successfully induced a sebogenic program, characterized by increased lipid accumulation and upregulation of PPARγ and FABP4, which was effectively attenuated by RTN and CPS. The 3D model successfully maintained essential compartmentalization, epithelial identity, and stromal organization. Furthermore, transcriptional analysis revealed that LA stimulation in 3D induced coordinated changes in extracellular matrix and basement membrane genes (including MMP13, MMP16, and THBS2), indicating that sebogenesis is directly coupled with structural remodeling and microenvironmental adaptation. Conclusion: model of a sebaceous gland organoid and established this platform as a relevant tool for mechanistic studies and for cosmetic screening of compounds targeting sebum regulation and sebaceous gland dysfunction.