Endocrine Abstracts

JOINT1024

Introduction: Dysregulation of adrenal steroidogenesis can cause imbalances in the secretion of mineralocorticoids, glucocorticoids, and androgens, leading to various endocrine disorders. We have previously established and extensively validated a human fetal adrenal ex vivo culture model, but no comparable model exists for adult human adrenal tissue. Developing an adult adrenal tissue culture model could facilitate studies of de novo and altered adrenal steroidogenesis in normal adrenal glands and hormone-producing adenomas, providing valuable insights towards establishing clinically relevant functional models. Despite important species-specific differences between human and mouse adrenals, we initially aim to establish an adult mouse adrenal ex vivo tissue culture model.

Materials and methods: Three ex vivo culture techniques were evaluated using adult mouse adrenal tissue: hanging drop, porous membranes, and agarose gel fragments. Adrenal tissue was cultured for 24 hours, 48 hours, or 5 days. Following each culture period, tissue was fixed, paraffin-embedded, and assessed for morphology (hematoxylin and eosin staining), proliferation (BrdU incorporation), and apoptosis (cleaved PARP staining). Subsequently, adrenal tissue will be treated with ACTH (1 nM) or ketoconazole (1 μM) to stimulate or inhibit steroidogenesis, respectively and effects on steroidogenesis will be determined by analyzing the production of selected adrenal steroid metabolites in the collected culture media.

Results: Analysis of the three different culture approaches and culture periods are currently ongoing. The optimal culture method will be determined based on preservation of tissue morphology (including intact cortical zones), minimal apoptosis, sustained cell proliferation, and functional responses to steroidogenic stimulation and inhibition.

Conclusion: Preliminary findings suggest that ex vivo culture of adult mouse adrenal tissue is feasible. Ongoing analyses will identify the most effective culture approach, providing a critical framework for future investigations into adrenal steroidogenesis. This model represents a significant first step toward enabling studies on both normal and pathological adrenal function in adult adrenal gland tissue.