Comparative analysis of 3D culture methodologies in prostate cancer cells.

Three-dimensional (3D) cell culture models are increasingly utilized in cancer research to better replicate in vivo tumor microenvironments. This study examines the effects of different 3D scaffolding materials, including Matrigel, GelTrex, and the plant-based GrowDex, on prostate cancer cell lines, with a particular emphasis on neuroendocrine prostate cancer (NEPC). Four cell lines (LNCaP, LASCPC-01, PC-3, and KUCaP13) were cultured in these scaffolds using the sandwich method to evaluate spheroid formation, cell viability, and gene expression. The results revealed that while all scaffolds supported cell viability, spheroid formation varied significantly: Matrigel promoted the most consistent spheroid formation, exhibiting the best results for LASCPC-01 among the tested scaffolds, though spheroid formation for LASCPC-01 was generally more limited than for LNCaP. Gene expression analysis indicated a reduction in androgen receptor (AR) expression in LNCaP cells across all scaffolds using the sandwich method. However, further experiments using a mini-dome method revealed that the expression of AR signaling genes and neuroendocrine marker genes varied depending on the scaffolds and culture methods. These findings highlight the scaffold-dependent variability in 3D culture outcomes and emphasize the need for standardized methodologies to ensure consistency and relevance in prostate cancer research.