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Abstract：
Craniofacial anomalies (CFA) are a diverse group of deformities, which affect the growth of the head and face. Dysregulation of cranial neural crest cell (NCC) migration, proliferation, differentiation, and/or cell fate specification have been reported to contribute to CFA. Understanding of the mechanisms through which cranial NCCs contribute for craniofacial development may lead to identifying meaningful clinical targets for the prevention and treatment of CFA. Isolation and culture of cranial NCCs in vitro facilitates screening and analyses of molecular cellular mechanisms of cranial NCCs implicated in craniofacial development. Here, we present a method for the isolation and culture of cranial NCCs harvested from the first branchial arch at early embryonic stages. Morphology of isolated cranial NCCs was similar to O9-1 cells, a cell line for neural crest stem cells. Moreover, cranial NCCs isolated from a transgenic mouse line with enhanced bone morphogenetic protein (BMP) signaling in NCCs showed an increase in their chondrogenic differentiation capacity, suggesting maintenance of their in vivo differentiation potentials observed in vitro. Taken together, our established method is useful to visualize cellular behaviors of cranial NCCs.
摘要：
颅面畸形（CFA）是一组不同的畸形，影响头部和面部的生长。颅神经c细胞（NCC）迁移的失调，扩散,分化,和/或细胞命运规范已被报道有助于CFA。了解颅骨NCCs促进颅面发育的机制可能会导致确定预防和治疗CFA的有意义的临床目标。体外分离和培养颅骨NCCs有助于筛选和分析与颅面发育有关的颅骨NCCs的分子细胞机制。这里,我们提出了一种在胚胎早期从第一分支弓收获的颅骨NCCs的分离和培养方法。分离的颅骨NCCs的形态与O9-1细胞相似，神经嵴干细胞的细胞系。此外，从NCC中具有增强的骨形态发生蛋白（BMP）信号传导的转基因小鼠系中分离出的颅骨NCC显示其软骨分化能力增加，表明维持其在体外观察到的体内分化潜能。一起来看,我们建立的方法可用于可视化颅骨NCCs的细胞行为。
