Related Experiment Video
Updated: Jul 30, 2025
08:57
3D Bioprinting of Murine Cortical Astrocytes for Engineering Neural-Like Tissue
Published on: July 16, 2021
6.6K
Protocol for 3D Bioprinting Mesenchymal Stem Cell-derived Neural Tissues Using a Fibrin-based Bioink
Milena Restan Perez1, Nadia Z Masri2, Jonathan Walters-Shumka2
1Axolotl Biosciences, 3800 Finnerty Road, Victoria, BC, V8W 2Y2, Canada.
Bio-Protocol
|May 15, 2023
Summary
This study details a comprehensive protocol for 3D bioprinting mesenchymal stem cells (MSCs) into functional dopaminergic neurons, bridging pre-printing cell culturing with post-printing analysis for disease modeling.
Area of Science:
- Regenerative Medicine
- Biotechnology
- Neuroscience
Background:
- Three-dimensional (3D) bioprinting creates living tissue models using cells and bioink.
- Stem cells, particularly mesenchymal stem cells (MSCs), are valuable for disease research and personalized medicine due to their regenerative and differentiation capabilities.
- Existing protocols for MSC bioprinting and cell culturing are often separate, creating a need for integrated methods.
Purpose of the Study:
- To provide a detailed, combined protocol for culturing, 3D bioprinting, and differentiating MSCs into dopaminergic neurons.
- To establish a reproducible method for generating patient-specific cell-based disease models.
- To facilitate research into degenerative diseases and potential therapeutic strategies.
Main Methods:
- Culturing of MSCs for 3D bioprinting.
- Preparation of Axolotl Biosciences TissuePrint bioinks (High Viscosity and Low Viscosity) and incorporation of MSCs.
- Utilizing BIO X and Aspect RX1 bioprinters with specified computer-aided design (CAD) files.
- Differentiation of 2D and 3D MSC cultures to dopaminergic neurons, including media preparation.
- Post-printing analysis including viability assays, immunocytochemistry, electrophysiology, and dopamine enzyme-linked immunosorbent assay (ELISA).
Main Results:
- A complete protocol for 3D bioprinting MSCs is presented, integrating cell culturing and bioprinting processes.
- Successful differentiation of bioprinted MSCs into dopaminergic neurons was achieved.
- Established methods for comprehensive post-printing analysis, including functional and biochemical assessments.
Conclusions:
- This integrated protocol enables the generation of 3D bioprinted MSC-derived dopaminergic neurons for disease modeling.
- The methodology supports personalized medicine approaches for studying neurodegenerative diseases.
- The protocol provides a foundation for advancing stem cell-based therapies and tissue engineering.
