Background: The human adrenal gland is composed of the cortex and the medulla, which contain different function cells. The aim of this study was to build a 3D culture system for human adrenal glands.
Methods: Human fetal adrenal tissues were digested into a cell suspension culture and processed in three-phase 3D cultures.
Results: Apparent spheroids could be seen from the 4th day on. After 21 days of 3D culture, steroid synthesis cells were evident via CYP17A1+ immunohistochemical staining and flow cytometry analysis. Electron microscopy analysis showed that these cells were present in lipid droplets in the cytoplasm. Meanwhile, TH+ cells represented catecholamine-producing cells, and these cells exhibited electron density particle gathering in the cytoplasm. Dehydroepiandrosterone and epinephrine syntheses were further confirmed via enzyme-linked immunosorbent assay.
Conclusions: We established a 3D culture system for human adrenal glands by using a sequential processing medium to facilitate cortical-medullary cell development.