Department of Craniofacial Sciences, School of Dental Medicine, University of Connecticut Health Center, Farmington, CT, USA.
Bone. 2010 Nov;47(5):948-58. doi: 10.1016/j.bone.2010.08.009. Epub 2010 Aug 20.
Transgenic mouse lines in which GFP expression is under the control of tissue- and stage specific promoters have provided powerful experimental tools for identification and isolation of cells at specific stage of differentiation along a lineage. In the present study, we used primary cell cultures derived from the dental pulp from pOBCol3.6GFP and pOBCol2.3GFP transgenic mice as a model to develop markers for early stages of odontoblast differentiation from progenitor cells. We analyzed the temporal and spatial expression of 2.3-GFP and 3.6-GFP during in vitro mineralization. Using FACS to separate cells based on GFP expression, we obtained relatively homogenous subpopulations of cells and analyzed their dentinogenic potentials and their progression into odontoblasts. Our observations showed that these transgenes were activated before the onset of matrix deposition and in cells at different stages of polarization. The 3.6-GFP transgene was activated in cells in early stages of polarization, whereas the 2.3-GFP transgene was activated at a later stage of polarization just before or at the time of formation of secretory odontoblast.
转绿色荧光蛋白（GFP）表达受组织和阶段特异性启动子控制的转基因小鼠系为鉴定和分离沿谱系分化的特定阶段的细胞提供了强大的实验工具。在本研究中，我们使用源自 pOBCol3.6GFP 和 pOBCol2.3GFP 转基因小鼠牙髓的原代细胞培养物作为模型，开发用于祖细胞向早期成牙本质细胞分化的标记物。我们分析了 2.3-GFP 和 3.6-GFP 在体外矿化过程中的时空表达。通过使用流式细胞术根据 GFP 表达分离细胞，我们获得了相对同质的细胞亚群，并分析了它们的牙本质发生潜力及其向成牙本质细胞的分化。我们的观察结果表明，这些转基因在基质沉积开始之前和在极化的不同阶段的细胞中被激活。3.6-GFP 转基因在早期极化阶段的细胞中被激活，而 2.3-GFP 转基因在极化后期被激活，就在分泌性成牙本质细胞形成之前或形成时。
