Shape, shrink, spheroid: a DIY high-throughput spheroid generation device

Area of Science:

• Biotechnology
• Cell Biology
• Drug Discovery

Background:

• 3D spheroids mimic in vivo conditions better than 2D cultures for drug testing and cancer research.
• Generating uniform spheroids at high throughput is challenging with current methods like ultra-low attachment plates or expensive microfabrication.

Purpose of the Study:

• To develop a low-cost, DIY device for high-throughput generation of uniform-sized 3D spheroids.
• To provide an accessible solution for resource-constrained laboratories needing advanced cell culture platforms.

Main Methods:

• A do-it-yourself (DIY) device using polydimethylsiloxane (PDMS) and agarose was created.
• Two variants (S1 and S2) were designed to fit 6-well and 12-well plates, generating 600 and 1200 spheroids, respectively.
• The device was validated with various cell lines and used for drug efficacy testing (IC50 estimation).

Main Results:

• The DIY device successfully generated hundreds of uniform-sized spheroids with minimal user intervention.
• Drug testing using the device yielded IC50 values comparable to conventional ultra-low attachment (ULA) plates.
• The device supports long-term culture via an optional perfusion system and can create size gradients for complex cell culture models.

Conclusions:

• A cost-effective, DIY method for high-throughput uniform spheroid generation is demonstrated.
• This technology democratizes advanced 3D cell culture for drug screening and biological research in labs with limited resources.
• The device's versatility extends to long-term culture and creating size-gradient spheroids for mimicking disease architecture.