Craniofacial cartilage organoids from human neural crest stem cells

We hypothesize that knowledge of cell signaling pathways that drive human neural crest differentiation into craniofacial chondrocytes may be obtained using human stem cells that self-organize into organoids. 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.本研究提出假说：可借助可自组织形成类器官（organoids）的人类干细胞，获取调控人类神经嵴向颅面软骨细胞分化的细胞信号通路相关知识。本研究开发了一套分化方案，可从人类胚胎干细胞来源的神经嵴干细胞中生成自组织的颅面软骨类器官。对软骨类器官进行组织学染色后，可见其具备弹性软骨典型的组织结构与染色特征。蛋白质及翻译后修饰（post-translational modification, PTM）质谱分析与单细胞核RNA测序（snRNA-seq）数据显示，软骨细胞类器官可高水平表达软骨细胞外基质（extracellular matrix, ECM）组分：包括多种胶原蛋白（collagens）、聚集蛋白聚糖（aggrecan）、基底膜蛋白聚糖（perlecan）、蛋白聚糖（proteoglycans）及弹性纤维（elastic fibers）。