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Fibrous polyisocyanide hydrogels for 3D cell culture applications
Hongbo Yuan1,2, Kaizheng Liu3, Melissa J J van Velthoven4
1Innovation and Research Institute of Hebei University of Technology in Shijiazhuang, Shijiazhuang, China. hongbo.yuan@kuleuven.be.
Nature Protocols
|May 30, 2025
Summary
Polyisocyanide (PIC) hydrogels offer a tunable, reproducible, and easy-to-handle synthetic alternative for 3D cell culture. Their biomimetic properties and thermoresponsive nature advance tissue engineering and disease modeling.
Area of Science:
- Biomaterials Science
- Cell Biology
- Tissue Engineering
Background:
- Three-dimensional (3D) cell culture models are crucial for tissue engineering, mechanobiology, disease modeling, and drug screening.
- Existing synthetic hydrogels often fail to mimic the extracellular matrix (ECM) effectively or present challenges in gel formation and cell extraction.
- The ECM's complexity is vital for cell development and function, extending beyond mere mechanical support.
Purpose of the Study:
- To introduce and provide protocols for polyisocyanide (PIC) hydrogels as advanced 3D cell culture materials.
- To highlight the biomimetic physical properties and ease of use of PIC hydrogels.
- To demonstrate the utility of PIC hydrogels across various cell types and experimental models.
Main Methods:
- Development of polyisocyanide (PIC) polymers for hydrogel formation.
- Characterization of PIC hydrogel fibrous architecture and mechanical properties.
- Utilization of thermoresponsive properties for rapid gelation at 37 °C and easy cell extraction at 5 °C.
- Demonstration of cell encapsulation, culture of various cell lines, primary cells, and organoids within PIC hydrogels.
- Inclusion of protocols for material preparation, cell encapsulation, and downstream analysis.
Main Results:
- PIC hydrogels exhibit a fibrous architecture and mechanical behavior that closely mimics natural biogels like collagen and fibrin.
- PIC hydrogels are fully synthetic, offering high tailorability and reproducibility.
- The thermoresponsive nature allows for instant gel formation at 37 °C and straightforward cell extraction upon cooling to 5 °C.
- PIC gels have been successfully applied in various cell culture applications, including organoid formation and in vivo experiments.
Conclusions:
- PIC hydrogels represent a significant advancement in 3D cell culture technology, overcoming limitations of existing synthetic materials.
- Their biomimetic properties, ease of handling, and versatility make them ideal for diverse applications in regenerative medicine and biological research.
- The provided protocols facilitate the adoption of PIC hydrogels by researchers in chemistry and cell biology laboratories.
