Ferreira Maidy Rehder Wimmers, Dernowsek Janaína, Passos Geraldo A, Bombonato-Prado Karina Fittipaldi
Cell Culture Laboratory - Department of Morphology, Stomatology and Physiology, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
Molecular Immunogenetics Group - Department of Genetics, Faculty of Medicine, University of São Paulo, Ribeirão Preto, SP, Brazil.
Arch Oral Biol. 2015 Apr;60(4):593-9. doi: 10.1016/j.archoralbio.2014.09.015. Epub 2014 Oct 19.
Expression of a large number of genes during differentiation of undifferentiated pulp cells into odontoblastic cells is still unknown, hence the aim of this investigation was to compare undifferentiated pulp cells (OD-21) and odontoblast-like cells (MDPC-23) through the assessment of cell stimulation and gene expression profiling.
The cells were cultured and after the experimental periods, there were evaluated cell proliferation and viability as well as alkaline phosphatase activity (ALP) and mineralization nodules. To evaluate gene expression it was used fluorescence cDNA microarray technology in addition to bioinformatics programmes such as SAM (significance analysis of microarrays). Gene expression was validated by Real Time PCR (qPCR).
The results showed that viability was above 80% in both cells, cell proliferation and ALP activity was higher in MDPC-23 cells and mineralization nodules were present only in the cultures of odontoblast-like cells. There were observed genes associated to odontogenesis with similar behaviour in both cell types, such as Il10, Traf6, Lef1 and Hspa8. Regions of the heatmap showed differences in induction and repression of genes such as Jak2 and Fas.
OD-21 cells share many genes with similar behaviour to MDPC-23 cells, suggesting their potential to differentiate into odontoblasts.
未分化牙髓细胞向成牙本质细胞分化过程中大量基因的表达情况仍不清楚，因此本研究的目的是通过评估细胞刺激和基因表达谱来比较未分化牙髓细胞（OD - 21）和成牙本质样细胞（MDPC - 23）。
培养细胞，在实验期结束后，评估细胞增殖、活力以及碱性磷酸酶活性（ALP）和矿化结节。为评估基因表达，除了使用生物信息学程序如SAM（微阵列显著性分析）外，还采用了荧光cDNA微阵列技术。基因表达通过实时PCR（qPCR）进行验证。
结果显示两种细胞的活力均高于80%，MDPC - 23细胞的细胞增殖和ALP活性更高，且矿化结节仅存在于成牙本质样细胞培养物中。在两种细胞类型中观察到与牙发生相关的具有相似行为的基因，如Il10、Traf6、Lef1和Hspa8。热图区域显示Jak2和Fas等基因在诱导和抑制方面存在差异。
OD - 21细胞与MDPC - 23细胞具有许多行为相似的基因，表明它们具有分化为成牙本质细胞的潜力。
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