Craniofacial chondrogenesis in organoids from human stem cell-derived neural crest cells
Lauren Foltz,
Nagashree Avabhrath,
Jean-Marc Lanchy,
Tyler Levy,
Anthony Possemato,
Majd Ariss,
Bradley Peterson,
Mark Grimes
Affiliations
Lauren Foltz
Division of Biological Sciences, Center for Biomolecular Structure and Dynamics, Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT 59812, USA
Nagashree Avabhrath
Division of Biological Sciences, Center for Biomolecular Structure and Dynamics, Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT 59812, USA
Jean-Marc Lanchy
Division of Biological Sciences, Center for Biomolecular Structure and Dynamics, Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT 59812, USA
Tyler Levy
Cell Signaling Technology, Danvers, MA 01923, USA
Anthony Possemato
Cell Signaling Technology, Danvers, MA 01923, USA
Majd Ariss
Cell Signaling Technology, Danvers, MA 01923, USA
Bradley Peterson
Pathology Consultants of Western Montana, Missoula, MT, USA
Mark Grimes
Division of Biological Sciences, Center for Biomolecular Structure and Dynamics, Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT 59812, USA; Corresponding author
Summary: Knowledge of cell signaling pathways that drive human neural crest differentiation into craniofacial chondrocytes is incomplete, yet essential for using stem cells to regenerate craniomaxillofacial structures. To accelerate translational progress, we developed a differentiation protocol that generated self-organizing craniofacial cartilage organoids from human embryonic stem cell-derived neural crest stem cells. Histological staining of cartilage organoids revealed tissue architecture and staining typical of elastic cartilage. Protein and post-translational modification (PTM) mass spectrometry and snRNA-seq data showed that chondrocyte organoids expressed robust levels of cartilage extracellular matrix (ECM) components: many collagens, aggrecan, perlecan, proteoglycans, and elastic fibers. We identified two populations of chondroprogenitor cells, mesenchyme cells and nascent chondrocytes, and the growth factors involved in paracrine signaling between them. We show that ECM components secreted by chondrocytes not only create a structurally resilient matrix that defines cartilage, but also play a pivotal autocrine cell signaling role in determining chondrocyte fate.
