CULTURE AND CHARACTERIZATION OF DENTAL FOLLICLE CELLS FROM RAT MOLARS
CULTURE AND CHARACTERIZATION OF DENTAL FOLLICLE CELLS FROM RAT MOLARS
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DOI：
10.1007/bf00319370
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发表时间：
1992-03-01
影响因子：
3.6
通讯作者：
FAN, W
中科院分区：
文献类型：
--
作者：
WISE, GE;LIN, F;FAN, W
由于牙囊是有限萌出牙齿萌出所必需的，因此本研究的目的是确定是否可以在体外培养毛囊细胞。 为了实现这一目标，从 6-7 日龄大鼠的第一和第二下颌磨牙（釉质生成的分泌阶段）中解剖出牙囊和相关的牙釉质器官，然后在促进成纤维细胞生长（牙囊的主要细胞类型）的培养基中培养。 培养的细胞生长至汇合并在实验前保持3代。 培养的细胞呈成纤维细胞形状，有突起伸长，透射电镜显示它们含有丰富的粗面内质网，但不形成桥粒。 抗波形蛋白的免疫荧光染色显示所有细胞均被染色，电子显微镜免疫金标记表明该抗体与中间丝相关。 SDS-聚丙烯酰胺凝胶电泳和蛋白质印迹显示，培养的细胞合成并分泌细胞外基质分子纤连蛋白和前胶原。 随后对透化和非透化细胞进行免疫荧光染色，证实细胞内和细胞外均存在纤连蛋白和 I 型胶原。 因此，基于所有上述特征，培养的细胞似乎是源自牙囊的成纤维细胞，尽管少数成纤维细胞可能源自插入牙槽骨和毛囊之间的未分化间充质细胞。 现在可以进行实验来确定这些培养细胞如何直接响应改变牙齿萌出率的生长因子。
Because the dental follicle is necessary for the eruption of teeth of limited eruption, it was the objective of this study to determine if the cells of the follicle could be cultured in vitro. To achieve this, dental follicles and associated enamel organs were dissected from the first and second mandibular molars of 6-7-day-old rats (secretory stage of amelogenesis), and then cultured in a medium that promotes fibroblast growth - the predominant cell type of the dental follicle. The cultured cells grew to confluency and were kept through 3 passages before experimentation. The cultured cells were fibroblastic in shape, elongate with processes, and transmission electron microscopy revealed that they contained an abundant rough endoplasmic reticulum, but did not form desmosomes. Immunofluorescent staining for anti-vimentin showed that all the cells stained and electron-microscopic immunogold labeling indicated that the antibody was associated with intermediate filaments. As revealed by SDS-polyacrylamide gel electrophoresis and Western blotting, the cultured cells synthesized and secreted the extracellular matrix molecules fibronectin and procollagens. Subsequent immunofluorescence staining of permeabilized and non-permeabilized cells confirmed the presence of fibronectin and type I collagen both intra- and extracellularly. Thus, based on all the above characteristics, the cultured cells appeared to be fibroblasts derived from the dental follicle, although a few of the fibroblasts may be derived from undifferentiated mesenchymal cells interposed between the alveolar bone and follicle. Experiments now can be conducted to determine how these cultured cells respond directly to growth factors that alter the rates of tooth eruption.