Three-Dimensional Bioprinting of Human iPSC-Derived Neuron-Astrocyte Cocultures for Screening Applications
September 29th, 2023
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.
Overview
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.
Key Study Components
Area of Science
- Neuroscience
- Cell Biology
- Biotechnology
Background
- 3D cell modeling has rapidly advanced, enabling more accurate representations of disease phenotypes.
- Traditional methods for 3D culture development are often labor-intensive and time-consuming.
- 3D bioprinting offers a solution by automating and scaling the development of complex cultures.
- This technology can produce numerous identical models rapidly, minimizing human error in the process.
Purpose of Study
- To create a high-throughput protocol for establishing 3D cocultures of neural cells.
- To improve the speed and convenience of model development in neuroscience research.
- To facilitate further investigation into the effects of 3D culture environments on neural cell types.
Methods Used
- The platform utilizes 3D bioprinting to create cocultures within hydrogel matrices.
- The biological model includes iPSC-derived neurons and astrocytes, grown in a controlled 3D environment.
- This protocol emphasizes efficient coculture establishment with minimal user intervention needed.
- Key timelines involve assessing cell viability and neurite outgrowth within a seven-day period.
- Hydrogel scaffolds are employed to support the structural integrity and functionality of the cells.
Main Results
- The coculture model exhibited high post-print cell viability and significant neurite outgrowth within seven days.
- Both cell types demonstrated the expression of maturity markers, indicating successful development.
- Rapid model creation supports more streamlined research applications and further testing.
- Conclusions highlight the potential of this protocol to overcome existing barriers in complex cell culture models.
Conclusions
- The study demonstrates a novel method for developing scalable 3D coculture systems in neuroscience research.
- This advancement may significantly impact the future of high-throughput assays in neural research.
- Insights gained from using 3D cultures could enhance understanding of neural mechanisms and plasticity.
