Induction of differentiation from human bone marrow-derived mesenchymal stem cell to odontoblast using epithelial-mesenchymal interactions

2010年，我重建了上皮细胞，形成了上皮细胞克隆。用双层共培养的方法将成釉细胞接种到间充质细胞上进行体外间充质相互作用。为此，我将牙源性形态发生相关的重组工具包基因Shh和Wnt基因导入间充质细胞，进行了类似的实验，分析了Shh和Wnt信号对成釉细胞的影响。2011年继续分析，用间充质细胞导入重组Shh和Wnt基因作为饲养细胞，成釉细胞上皮集落数明显增加，最大集落直径明显增大。此外，将成釉细胞接种到丝裂霉素C处理的间充质细胞上，培养2周后成釉细胞形成结节，Shh和Wnt信号组Notch1基因表达受到抑制，从而调节细胞分化。WNT信号完全抑制成釉细胞Runx2/Cbfa1基因的表达。今年，我又生产了胶原支架、间充质细胞和成釉细胞的混合物，并将其移植到裸鼠背部的皮下。我没有达到牙齿再生，因为移植的混合物发生了致癌变化。

In 2010, epithelial colonies were formed after I reconstructed epithelial. mesenchymal interactions in vitro by double-layer co-culture method that inoculated ameloblasts on mesenchymal cells. Therefore I introduced recombinant Shh and Wnt gene of tool kit genes these related odontogenic morphogenesis to mesenchymal cells and carried out similar experiment, analyzed influences those signals of Shh and Wnt gave adamant oblast.I continued analyses as for 2011, when using mesenchymal cells introduced recombinant Shh and Wnt gene as feeder cells, epithelial colony count of ameloblasts increased dominantly, and diameter of a colony at the maximum was enlarged significantly. In addition, ameloblasts formed the nodules after two weeks culture when ameloblasts were inoculated on mesenchymal cells treated mitomycin C. Nodules were bigger significantly, Notch1 gene expression was inhibited that regulated cell differentiation in Shh and Wnt signaling groups. Wnt signal completely inhibited Runx2/Cbfa1 gene expression of ameloblasts. More this year, I produced the mixture of collagen scaffold, mesenchymal cell and ameloblast, and transplanted the mixtures into back of nude mice subcutaneously. I do not reached tooth regeneration, because of oncogenic change of transplanted mixture.