Magnetically Bioprinted 3D Spheroid Co-cultures: A Method for Co-culturing Cancerous Cells and Fibroblasts

- In magnetic bioprinted 3D co-cultures, multiple cell types form 3D structures or spheroids under magnetic fields' influence. To set up the culture, first, prepare single-cell suspensions of cancer cells and fibroblasts in their appropriate medium. Next, add biocompatible magnetic nanoparticles to both suspensions. These nanomaterials contain positively charged poly L-lysine residues that facilitate the nonspecific binding of magnetic particles to the cell membrane via electrostatic interactions.

Incubate to achieve maximum magnetization of cancer cells and fibroblasts. Subsequently, mix both cell suspensions in the desired ratio. Transfer this cell-nanoparticle suspension to a cell-repellent culture plate, specifically optimized to prevent cell attachment. Mount this setup on a magnetic drive and incubate. Magnetic forces guide the aggregation or bioprinting of magnetized cancer cells and fibroblasts into 3D spheroid structures at the bottom of the well.

Once spheroids form, remove the magnet and allow the 3D co-cultures to grow to the desired volume. In this protocol, we will demonstrate the generation of three-dimensional co-culture spheroids from pancreatic cancer cells and PS