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Protocol for generating human craniofacial cartilage organoids from stem-cell-derived neural crest cells
Nagashree Avabhrath1, Lauren Foltz1, Mark Grimes1
1Division of Biological Sciences, Center for Biomolecular Structure and Dynamics, and Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT 59812, USA.
STAR Protocols
|December 21, 2024
Summary
This study details a protocol for creating craniofacial cartilage organoids from human stem cells. This method uses neural crest stem cells (NCSCs) to regenerate craniofacial structures and model diseases.
Area of Science:
- Biotechnology
- Developmental Biology
- Stem Cell Research
Background:
- Craniofacial cartilage regeneration is crucial for reconstructive surgery and understanding developmental disorders.
- Existing methods for generating craniofacial cartilage are limited in recapitulating in vivo development.
Purpose of the Study:
- To establish a robust protocol for generating craniofacial cartilage organoids from human pluripotent stem cells.
- To enable advanced craniofacial reconstruction and disease modeling through self-organizing organoids.
Main Methods:
- Induction of human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) into neural crest stem cells (NCSCs) using small molecules and growth factors.
- Isolation of NCSCs via magnetic bead sorting.
- Culturing NCSCs under specific conditions to promote self-organization into craniofacial cartilage organoids.
Main Results:
- Successful generation of craniofacial cartilage organoids from human stem cells.
- Demonstration of NCSC migration and self-organization into cartilage structures.
- Protocol provides a reproducible method for craniofacial chondrogenesis.
Conclusions:
- The developed protocol effectively generates craniofacial cartilage organoids, mimicking natural chondrogenesis.
- This advancement holds significant potential for craniofacial tissue engineering and disease pathology studies.
- Facilitates future research in craniofacial development and regenerative medicine.
