Jing Yan, Li Chaoyuan, Feng Jian Q
Department of Orthodontics, Texas A&M University College of Dentistry, Dallas, TX, USA.
Department of Biomedical Sciences, Texas A&M University College of Dentistry, Dallas, TX, USA.
Methods Mol Biol. 2019;1922:39-48. doi: 10.1007/978-1-4939-9012-2_5.
The cell lineage tracing system has been used predominantly in developmental biology studies. The Cre recombinase allows for the activation of the reporter in a specific cell line and all progeny. In this protocol, we will introduce how the cell lineage tracing technique can be performed in the investigation of dentinogenesis by using Gli1-Cre; R26R compound mice. Moreover, we combined cell lineage tracing in conjunction with immunofluorescence-to further define cell fate by analyzing the expression of specific cell markers for odontoblasts. This combination not only broadens the application of cell lineage tracing but also simplifies the generation of compound mice. More importantly, the number, location, and differentiation status of parent cell progeny can be displayed simultaneously, providing more information than cell lineage tracing or immunofluorescence alone. In conclusion, the co-application of cell lineage tracing technique and immunofluorescence is a powerful tool for investigating cell biology in the field of dentinogenesis and tooth development.
细胞谱系追踪系统主要用于发育生物学研究。Cre重组酶可使报告基因在特定细胞系及其所有子代细胞中激活。在本方案中，我们将介绍如何通过使用Gli1-Cre; R26R复合小鼠，在牙本质形成研究中进行细胞谱系追踪技术。此外，我们将细胞谱系追踪与免疫荧光相结合，通过分析成牙本质细胞特异性细胞标志物的表达来进一步确定细胞命运。这种结合不仅拓宽了细胞谱系追踪的应用范围，还简化了复合小鼠的制备。更重要的是，亲代细胞子代的数量、位置和分化状态可以同时显示，比单独使用细胞谱系追踪或免疫荧光提供更多信息。总之，细胞谱系追踪技术与免疫荧光的联合应用是牙本质形成和牙齿发育领域研究细胞生物学的有力工具。
