Adipose Tissue-Derived Stem Cell in Vitro Differentiation in a Three-Dimensional Dental Bud Structure
Adipose Tissue-Derived Stem Cell in Vitro Differentiation in a Three-Dimensional Dental Bud Structure
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DOI：
10.1016/j.ajpath.2011.01.055
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发表时间：
2011-05-01
影响因子：
6
通讯作者：
Curcio, Francesco
中科院分区：
文献类型：
--
作者：
Ferro, Federico;Spelat, Renza;Curcio, Francesco
牙齿的形态发生需要颅神经嵴来源的间充质细胞和气孔上皮之间的相互作用，调节牙齿的形态发生和分化。我们展示了间充质来源的单个干细胞群体如何在体外被诱导转分化成类似于牙芽的结构。脂肪组织中存在干细胞以前已有报道。我们将人脂肪组织来源的干细胞原代培养在诱导成牙培养基中，并在三维条件下培养聚集体。四周后，细胞形成了类似于牙芽的三维组织结构。通过RT-PCR、免疫印迹、MC和理化分析检测牙组织相关标志物的表达，测定谱系特异性mRNA和蛋白质。在诱导培养基中，细胞对成釉细胞和成牙本质细胞标志物的mRNA和蛋白质均呈阳性。此外，细胞表达上皮，间充质和基底膜标记的位置关系类似的生理牙齿形态发生。物理化学分析显示，200 nm和50 nm的定向羟基磷灰石晶体显示在体内的牙釉质和牙本质，分别。总之，我们表明，脂肪组织来源的干细胞在体外可以转分化产生一个特定的三维组织和表型类似的牙芽，即使在没有结构基质或支架，以指导发展过程。(Am J Pathol 2011，178：2299 2310; DOI：10.1016/j.ajpath.2011.01.055）
Tooth morphogenesis requires sequential and reciprocal interactions between the cranial neural crest derived mesenchymal cells and the stomadial epithelium, which regulate tooth morphogenesis and differentiation. We show how mesenchyme-derived single stem cell populations can be induced to transdifferentiate in vitro in a structure similar to a dental bud. The presence of stem cells in the adipose tissue has been previously reported. We incubated primary cultures of human adipose tissue derived stem cells in a dental-inducing medium and cultured the aggregates in three-dimensional conditions. Four weeks later, cells formed a three-dimensional organized structure similar to a dental bud. Expression of dental tissue related markers was tested assaying lineage-specific mRNA and proteins by RT-PCR, immunoblot, MC, and physical-chemical analysis. In the induction medium, cells were positive for ameloblastic and odontoblastic markers as both mRNAs and proteins. Also, cells expressed epithelial, mesenchymal, and basement membrane markers with a positional relationship similar to the physiologic dental morphogenesis. Physical-chemical analysis revealed 200-nm and 50-nm oriented hydroxyapatite crystals as displayed in vivo by enamel and dentin, respectively. In conclusion, we show that adipose tissue derived stem cells in vitro can transdifferentiate to produce a specific three-dimensional organization and phenotype resembling a dental bud even in the absence of structural matrix or scaffold to guide the developmental process. (Am J Pathol 2011, 178:2299 2310; DOI: 10.1016/j.ajpath.2011.01.055)