DOI: 10.3791/65856-v
This study presents a protocol for the efficient production of 3D-bioprinted cocultures of iPSC-derived neurons and astrocytes within hydrogel scaffolds. The developed model operates in 96- or 384-well formats and demonstrates high post-print viability and neurite outgrowth within seven days, while expressing maturity markers for both cell types. This approach aims to enhance the throughput and automation of 3D cell culture systems.
Here, we present a protocol to produce 3D-bioprinted cocultures of iPSC-derived neurons and astrocytes. This coculture model, generated within a hydrogel scaffold in 96- or 384-well formats, demonstrates high post-print viability and neurite outgrowth within 7 days and shows the expression of maturity markers for both cell types.
3D cell modeling is a novel field that has exponentially expanded in the past decade. These models have been shown to both facilitate neuronal growth and more accurately represent disease phenotypes. However, we believe there is a shift towards making these models higher throughput and a necessity to embrace automation within development.
Traditional methods of developing 3D cultures can be laborious and time-consuming to establish, but 3D bioprinting is a technology that can be applied to scale up these development processes. This technology allows for hundreds of identical models to be created efficiently and without human error. This protocol develops complex cultures because the neural cells are grown in 3D in biologically active hydrogel matrices.
But critically, this protocol prioritizes speed and convenience in model development, which can be lacking in this field and can hinder the implementation into industry. This protocol defines a method to establish many 3D cocultures very efficiently with limited input from users. We hope this will remove barriers to using complex cell culture models within high throughput assays and facilitate further investigation of the effect of 3D culture on neural cell types.
