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bioRxiv · 10.1101/2025.03.11.642589

Comparative Analysis of 3D Culture Methodologies in Prostate Cancer Cells

Foster, E.·Wardhana, O.·Zeng, Z.·Lu, X.
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Abstract

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 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 robust spheroids, especially for LASCPC-01, whereas GrowDex exhibited limitations for certain cell lines. Gene expression analysis indicated a consistent reduction in androgen receptor (AR) expression in LNCaP cells across all scaffolds, suggesting a potential shift towards a neuroendocrine phenotype. However, the expression of neuroendocrine markers varied depending on the scaffold and culture method, with the mini-domes method in Matrigel leading to decreased expression of both castration-resistant prostate cancer (CRPC) and NEPC markers. 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. HighlightsO_LIComparing of Matrigel, GelTrex, and GrowDex in 3D prostate cancer culture C_LIO_LIIdentifying scaffold-dependent spheroid formation and gene expression shifts C_LIO_LIObserving consistent AR reduction and variable neuroendocrine marker changes C_LIO_LIHighlighting the need for standard 3D culture methods in cancer studies C_LI

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Foster, E., Wardhana, O., Zeng, Z., Lu, X.. 2025-03-13. Comparative Analysis of 3D Culture Methodologies in Prostate Cancer Cells. https://doi.org/10.1101/2025.03.11.642589

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