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.

我们提出如下假说：可利用可自组织形成类器官（organoids）的人类干细胞，解析驱动人类神经嵴分化为颅面软骨细胞的细胞信号通路机制。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获得可自组织的颅面软骨类器官。对软骨类器官开展组织学染色后，可见其组织结构与弹性软骨的典型染色特征相符。蛋白质组与翻译后修饰（post-translational modification, PTM）质谱分析、单细胞核RNA测序（snRNA-seq）数据表明，软骨细胞类器官可显著高表达软骨细胞外基质（ECM）组分，涵盖多种胶原蛋白、聚集蛋白聚糖、串珠蛋白聚糖（perlecan）、蛋白聚糖及弹性纤维。