Odontoblast differentiation of human dental pulp cells in explant cultures
Odontoblast differentiation of human dental pulp cells in explant cultures
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DOI：
10.1007/pl00005833
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发表时间：
2000-02-01
影响因子：
4.2
通讯作者：
Magloire, H
中科院分区：
文献类型：
--
作者：
Couble, ML;Farges, JC;Magloire, H
为了阐明人牙本质形成的机制，我们建立了一套促进牙髓细胞向成牙本质细胞分化的细胞培养体系。人牙外植体在添加10%或15%胎牛血清的Eagle‘s基本培养液中培养，添加或不添加β-甘油磷酸(β-GP)。在培养的牙髓细胞中加入β-GP诱导成牙本质细胞的特征。细胞极化，其中一些细胞呈现典型的细胞伸展。在某些情况下，细胞与它们的突起朝向相同的方向，并形成类似于体内连接成牙本质细胞的末端网的连接复合体。精细的结构分析显示，成牙本质细胞小体存在典型的细胞内细胞器，而这个过程只包含细胞骨架元素和分泌囊泡。极化细胞沉积在塑料培养皿上，形成丰富而有组织的I型胶原基质，随后出现矿化区。X-射线微区分析表明，钙和磷的存在，电子衍射图证实了该矿物的磷灰石晶体结构。α1(1)型胶原基因在所有极化细胞中均有高表达，而牙本质涎蛋白基因主要在矿化区表达。这种细胞培养体系允许牙髓细胞在形态和功能水平上分化为成牙本质细胞。此外，这些细胞呈现出类似于成牙本质细胞层的空间组织。
In order to elucidate the mechanisms involved in human dentin formation, we developed a cell culture system to promote differentiation of dental pulp cells into odontoblasts. Explants from human teeth were cultured in Eagle's basal medium supplemented with 10% or 15% fetal calf serum, with or without beta-glycerophosphate (beta GP). Addition of beta GP to the culture medium induced odontoblast features in the cultured pulp cells. Cells polarized and some of them exhibited a typical cellular extension. In some cases, cells aligned with their processes oriented in the same direction and developed junctional complexes similar to the terminal web linking odontoblasts in vivo. Fine structural analyses showed the presence of typical intracellular organelles of the odontoblast body, whereas the process contained only cytoskeleton elements and secretory vesicles. Polarized cells deposited onto the plastic dishes an abundant and organized type I collagen-rich matrix with areas of mineralization appearing thereafter. X-ray microanalysis showed the presence of calcium and phosphorus and the electron diffraction pattern confirmed the apatitic crystal structure of the mineral. High expression of alpha 1(1) collagen mRNAs was detected in all polarized cells whereas dentin sialoprotein gene was mainly expressed in mineralizing areas. This cell culture system allowed for the differentiation of pulp cells into odontoblasts, at both the morphological and functional level. Moreover, these cells presented a spatial organization similar to the odontoblastic layer.