Multi-Inlet Spheroid Generator for High-Throughput Combinatorial Drug Screening Based on the Tumor Microenvironment

Area of Science:

• Biotechnology
• Cancer Research
• Drug Discovery

Background:

• Tumor spheroids are crucial for drug screening and understanding tumor physiology.
• The hanging drop method is ideal for high-throughput screening (HTS) but lacks sufficient liquid-holding capacity.
• Increased pressure from added substances can cause hanging drops to detach, limiting current methods.

Purpose of the Study:

• To develop a novel spheroid formation method that overcomes the limitations of existing hanging drop techniques.
• To enhance the stability and liquid-holding capacity of tumor spheroids for advanced drug screening.
• To create a versatile platform for both high-throughput anticancer drug screening and recapitulating the tumor microenvironment (TME).

Main Methods:

• Introduction of a multi-inlet spheroid generator (MSG) for stable liquid addition into spheroids.
• Utilizing side inlets to introduce drugs or cells without increasing applied force on the hanging drop.
• Controlling the volume and sequence of liquid injections via adjustable side inlet diameters and multiple inlets.

Main Results:

• The MSG successfully enabled stable addition of liquids through side inlets, maintaining spheroid integrity.
• Liquid volume and injection sequences were precisely controlled, demonstrating the system's versatility.
• The MSG platform showed feasibility in clinical applications, including testing drug efficacy on patient-derived cancer (PDC) cells and manipulating the TME composition.

Conclusions:

• The multi-inlet spheroid generator (MSG) offers a stable and controllable method for tumor spheroid manipulation.
• This technology significantly advances high-throughput screening (HTS) of anticancer drugs.
• The MSG platform provides a powerful tool for accurately recapitulating the tumor microenvironment (TME) for research and clinical applications.