2. Academic Validation
  3. Protocol for generating human craniofacial cartilage organoids from stem-cell-derived neural crest cells

Protocol for generating human craniofacial cartilage organoids from stem-cell-derived neural crest cells

  • STAR Protoc. 2025 Mar 21;6(1):103532. doi: 10.1016/j.xpro.2024.103532.
Nagashree Avabhrath 1,  Lauren Foltz 2,  Mark Grimes 3
Affiliations

Affiliations

  • 1 Division of Biological Sciences, Center for Biomolecular Structure and Dynamics, and Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT 59812, USA. Electronic address: nagashree.avabhrath@umconnect.umt.edu.
  • 2 Division of Biological Sciences, Center for Biomolecular Structure and Dynamics, and Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT 59812, USA.
  • 3 Division of Biological Sciences, Center for Biomolecular Structure and Dynamics, and Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT 59812, USA. Electronic address: mark.grimes@mso.umt.edu.
PMID: 39708322 DOI: 10.1016/j.xpro.2024.103532
Abstract

Here, we present a protocol to generate craniofacial cartilage organoids from human stem cells via neural crest stem cells (NCSCs). We describe steps for inducing human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) to form NCSCs using sequential treatments of small molecules and growth factors and isolating NCSCs by magnetic bead sorting. We then detail procedures for defining conditions where NCSCs migrate together and self-organize into craniofacial cartilage organoids. Recapitulating craniofacial chondrogenesis will facilitate craniofacial reconstruction and disease modeling. For complete details on the use and execution of this protocol, please refer to Foltz et al.1.

Keywords

cell differentiation; classification description: stem cells; organoids.

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