Visakan Gayathri, Su Jingtan, Moradian-Oldak Janet
Center for Craniofacial Molecular Biology, Herman Ostrow School of Dentistry, University of Southern California, Los Angeles, CA, United States.
Data Brief. 2022 May 5;42:108233. doi: 10.1016/j.dib.2022.108233. eCollection 2022 Jun.
This article contains data related to the research article in this issue titled ameloblastin promotes polarization of ameloblast cell lines in a 3D cell culture system (Visakan et al., 2022). In the process of amelogenesis, the organic matrix components are pivotal to the establishment of ameloblast-matrix adhesion. Here we employ immortalized ameloblast cell lines and analyse their morphological changes in 3D cell culture when cultured in the presence of recombinant enamel matrix proteins- ameloblastin and amelogenin compared with controls. The recombinant proteins that were purified using high-performance liquid chromatography (HPLC) were characterized using SDS-gel electrophoresis. A 3D-on-top culture technique was employed, and the cells were analysed 24 and 72 h post inoculation using fluorescent and confocal microscopy for qualitative and quantitative changes. Aspect ratio of cells was measured and used as the parameter to compare between test proteins and controls. Repeated measurements of aspect ratio were recorded to analyse for statistical significance. Additionally, three distinct gel substrates were studied to examine the effect of composition and stiffness of the substrate on cell behaviour. The cells in the 3D culture were fixed and labelled using antibodies to junctional complex, polarity and tight junctional proteins following protocols for whole culture fixation. Co-localization between membrane and specific antibody labels were examined under confocal microscopy.
本文包含与本期研究论文相关的数据，该论文题为《成釉蛋白在三维细胞培养系统中促进成釉细胞系的极化》（维萨坎等人，2022年）。在釉质形成过程中，有机基质成分对于成釉细胞与基质黏附的建立至关重要。在此，我们使用永生化成釉细胞系，并分析与对照组相比，在重组釉质基质蛋白——成釉蛋白和釉原蛋白存在的情况下，其在三维细胞培养中的形态变化。使用高效液相色谱（HPLC）纯化的重组蛋白通过SDS凝胶电泳进行表征。采用三维顶层培养技术，并在接种后24小时和72小时使用荧光显微镜和共聚焦显微镜对细胞进行定性和定量变化分析。测量细胞的纵横比，并将其用作比较测试蛋白与对照组的参数。记录纵横比的重复测量值以分析统计学意义。此外，研究了三种不同的凝胶底物，以检查底物的组成和硬度对细胞行为的影响。按照全培养固定方案，使用针对连接复合体、极性和紧密连接蛋白的抗体对三维培养中的细胞进行固定和标记。在共聚焦显微镜下检查膜与特异性抗体标记之间的共定位。
