Optimization of OSCC spheroid generation for high-throughput drug screening

Area of Science:

• Oncology
• Biotechnology
• Cancer Research

Background:

• Three-dimensional (3D) spheroid models more accurately mimic the in vivo tumor microenvironment compared to traditional two-dimensional (2D) cultures.
• This makes 3D spheroids valuable tools for preclinical drug screening in oral squamous cell carcinoma (OSCC).

Purpose of the Study:

• To establish and characterize scaffold-free 3D spheroid models for OSCC using different culture methods.
• To compare the drug response of 3D spheroids with 2D cultures for OSCC cell lines.

Main Methods:

• Scaffold-free spheroids were generated from CAL 27 and OECM-1 OSCC cell lines using methylcellulose suspension, agarose-coated plates, and ultra-low attachment (ULA) plates.
• Morphology, uniformity, viability, necrosis, gene expression of cancer hallmark genes, and drug response to cisplatin and doxorubicin were assessed.

Main Results:

• Compact, uniform spheroids formed within 72 hours, with ULA plates yielding the highest reproducibility and structural integrity.
• 3D spheroids exhibited central necrosis, modulated cancer hallmark gene expression (hypoxia, angiogenesis, EMT), and significantly higher resistance to cisplatin and doxorubicin compared to 2D cultures.

Conclusions:

• ULA-based 3D spheroids provide a reproducible, stable, and scalable model that accurately reflects OSCC tumor biology and chemoresistance.
• These 3D models are suitable for high-throughput drug screening and advancing translational cancer research in OSCC.