Development and Characterization of 3-Dimensional Cell Culture Models of Adrenocortical Carcinoma

Endocrinology. 2024 Nov 26;166(1):bqae159. doi: 10.1210/endocr/bqae159.

Abstract

Adrenocortical carcinoma (ACC) is a rare malignancy of the adrenal cortex that is associated with a poor prognosis. Developing effective treatment options for ACC is challenging owing to the current lack of representative preclinical models. This study addressed this limitation by developing and characterizing 3-dimensional (3D) cell cultures incorporating the ACC cell lines, MUC-1, HAC15, and H295R in a type I collagen matrix. ACC tissue samples were analyzed by immunohistochemistry to determine the presence of type I collagen in the tumor microenvironment. Cell viability and proliferation were assessed using flow cytometry and confocal microscopy. mRNA expression of steroidogenic enzymes and steroid secretion was analyzed by comparing the 3D and monolayer cell culture models. All cells were successfully cultured in a type I collagen matrix, which is highly expressed in the ACC tumor microenvironment and showed optimal viability until day 7. All 3 models showed increased metabolic and proliferative activity over time. Three-dimensional cell cultures were steroidogenic and demonstrated increased resistance to the gold standard chemotherapy, mitotane, compared with monolayer. The use of these models may lead to an improved understanding of disease pathology and provide a better representative platform for testing and screening of potential therapies.

Keywords: 3D cell culture models; adrenocortical carcinoma; drug resistance; preclinical models.

MeSH terms

  • Adrenal Cortex Neoplasms* / genetics
  • Adrenal Cortex Neoplasms* / metabolism
  • Adrenal Cortex Neoplasms* / pathology
  • Adrenocortical Carcinoma* / genetics
  • Adrenocortical Carcinoma* / metabolism
  • Adrenocortical Carcinoma* / pathology
  • Cell Culture Techniques / methods
  • Cell Culture Techniques, Three Dimensional* / methods
  • Cell Line, Tumor
  • Cell Proliferation*
  • Cell Survival*
  • Collagen Type I / genetics
  • Collagen Type I / metabolism
  • Humans
  • Mitotane / pharmacology
  • Tumor Microenvironment

Substances

  • Mitotane
  • Collagen Type I