Hard palate, developed by embryo neural crest stem cells, is a tissue with strong regenerative abilities. It is considered an abundant source of progenitor cells, forming various mesenchymal tissues. Rabbits are more suitable models than murine animals for regenerative preclinical study of the head and neck, owing to their larger size. However, there are no reports of the existence or characteristics of neural crest stem cells in the hard palate of rabbits. In this study, we demonstrate for the first time the presence of nestin-, Sox2-, and p75-positive neural crest stem cells obtained from the hard palate of rabbits and the properties of these cells. Flow cytometry analysis revealed that CD29, CD44, and CD81 were positive; and CD11b, CD34, and CD90 were negative on the ex vivo expanded palatal progenitor cells. Finally, we differentiated them into cells of mesenchymal lineages (bone, cartilage, and fat) in vitro, and in three-dimensional fabricated polycaprolactone and polycaprolactone–tricalcium phosphate scaffolds. Taken together, our data showed the existence of rabbit palatum-derived mesenchymal progenitor cells, and successful fabrication of progenitor cell-loaded biodegradable scaffold using three-dimensional printing. This study will open avenues for new tissue engineering strategies for cell therapy using three-dimensional printing with scaffolds for reconstruction of head and neck defects.

中文翻译：

---

兔腭源间充质祖细胞在 2D 基质和 3D 打印结构上的三系分化

硬腭由胚胎神经嵴干细胞发育而成，是一种再生能力强的组织。它被认为是祖细胞的丰富来源，可形成各种间充质组织。兔子比鼠类动物更适合用于头部和颈部的再生临床前研究，因为它们的体型较大。然而，没有关于兔硬腭中神经嵴干细胞的存在或特征的报道。在这项研究中，我们首次证明了从兔硬腭中获得的巢蛋白、Sox2 和 p75 阳性神经嵴干细胞的存在以及这些细胞的特性。流式细胞术分析显示CD29、CD44、CD81阳性；CD11b、CD34 和 CD90 在体外扩增的腭祖细胞中呈阴性。最后，我们在体外和三维制造的聚己内酯和聚己内酯-磷酸三钙支架中将它们分化为间充质谱系（骨、软骨和脂肪）细胞。综上所述，我们的数据表明存在兔腭源间充质祖细胞，并使用 3D 打印成功制造了加载祖细胞的可生物降解支架。这项研究将为细胞治疗的新组织工程策略开辟道路，使用带有支架的 3D 打印重建头颈部缺陷。我们的数据显示存在兔腭源间充质祖细胞，并使用 3D 打印成功制造了加载祖细胞的可生物降解支架。这项研究将为细胞治疗的新组织工程策略开辟道路，使用带有支架的 3D 打印重建头颈部缺陷。我们的数据显示存在兔腭源间充质祖细胞，并使用 3D 打印成功制造了加载祖细胞的可生物降解支架。这项研究将为细胞治疗的新组织工程策略开辟道路，使用带有支架的 3D 打印重建头颈部缺陷。