Tamaki Yuichi, Nakahara Taka, Ishikawa Hiroshi, Sato Soh
Department of Developmental and Regenerative Dentistry, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan.
Odontology. 2013 Jul;101(2):121-32. doi: 10.1007/s10266-012-0075-0. Epub 2012 Jul 7.
Mesenchymal stem cells derived from human teeth and bone marrow have been characterized by many research groups, but demonstrate inconsistent cellular phenotypes or functions, partly because of differences in culture methodology. Therefore, our aims were to resolve these inconsistencies and discuss the potential uses of these cells in research/clinical applications. We isolated and characterized dental stem cells (DSCs) from the dental pulp, periodontal ligament, apical papilla (APSCs) and dental follicle (DFSCs) of mature and immature teeth, along with bone marrow-derived stem cells (BMSCs) from the iliac crest. We compared the clonogenic and proliferative potentials of these cells in terms of colony-forming efficiency, proliferation potential, population doubling time and cell cycle. All DSCs, particularly APSCs and DFSCs, possessed greater proliferative potential than BMSCs. All stem cells expressed typical mesenchymal and embryonic markers, and developed alizarin red-positive mineralization nodules and Oil red O-positive lipid droplets when cultured in osteogenic and adipogenic media, respectively. Immunocytochemistry revealed that all stem cells developed neuronal markers when cultured in a control medium without neural inductive supplements. After 7 days of neurogenic culture, the differentiated cells showed a transition from fibroblast-like to neuron-like cell bodies with long processes, suggesting that the stem cells differentiated into mature neurons. Karyotyping confirmed that the stem cells maintained a normal karyotype and were chromosomally stable. Our results provide new insights into the physiological properties of stem cells with a normal karyotype and indicate that DSCs are appropriate for basic research and clinical applications.
许多研究小组已对源自人牙齿和骨髓的间充质干细胞进行了表征，但这些细胞表现出不一致的细胞表型或功能，部分原因是培养方法存在差异。因此，我们的目标是解决这些不一致性问题，并讨论这些细胞在研究/临床应用中的潜在用途。我们从成熟和未成熟牙齿的牙髓、牙周韧带、根尖乳头（APSCs）和牙囊（DFSCs）中分离并表征了牙干细胞（DSCs），同时还从髂嵴中分离了骨髓来源的干细胞（BMSCs）。我们从集落形成效率、增殖潜力、群体倍增时间和细胞周期等方面比较了这些细胞的克隆形成和增殖潜力。所有DSCs，尤其是APSCs和DFSCs，比BMSCs具有更强的增殖潜力。所有干细胞均表达典型的间充质和胚胎标志物，并且分别在成骨和成脂培养基中培养时会形成茜素红阳性矿化结节和油红O阳性脂滴。免疫细胞化学显示，在没有神经诱导添加剂的对照培养基中培养时，所有干细胞均会形成神经元标志物。经过7天的神经源性培养后，分化的细胞显示出从成纤维细胞样细胞体向具有长突起的神经元样细胞体的转变，这表明干细胞分化为成熟神经元。核型分析证实干细胞保持正常核型且染色体稳定。我们的结果为具有正常核型的干细胞的生理特性提供了新见解，并表明DSCs适用于基础研究和临床应用。
