Flipped Well-Plate Hanging-Drop Technique for Growing Three-Dimensional Tumors

Area of Science:

• Biotechnology
• Cancer Research
• Cell Biology

Background:

• Three-dimensional (3D) tumor cultures are valuable in vitro models for cancer research.
• Current 3D tumor culturing methods often lack standardization and are difficult to replicate outside of specialized labs.
• There is a need for accessible and user-friendly 3D tumor culture techniques.

Purpose of the Study:

• To introduce a straightforward and user-friendly 3D tumor culture method using standard 96-well plates.
• To demonstrate the applicability of this method for scaffold-free and scaffold-based tumor culture.
• To facilitate the standardization and broader adoption of 3D tumor models in research.

Main Methods:

• Development of the well-plate flip (WPF) method for generating hanging drops in standard 96-well plates.
• Utilizing human colorectal carcinoma cells (HCT116) for spheroid generation.
• Demonstration of both scaffold-free and matrix-assisted 3D tumor culture techniques.

Main Results:

• Successful generation of large, scaffold-free 3D tumor spheroids exceeding 1.5 mm in diameter.
• The WPF method is adaptable for various cell biology applications, including cancer research.
• Proof-of-concept for matrix-assisted tumor culture using the developed approach.

Conclusions:

• The WPF method offers a simple, accessible, and versatile approach for 3D tumor spheroid formation.
• This technique addresses the limitations of current 3D culture methods, promoting standardization.
• The WPF method has broad applicability in basic and applied biological and engineering research, advancing cancer modeling.