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

2010年，我重建上皮后，上皮菌落形成。用双层共培养法在间充质细胞上接种成釉细胞，观察间充质细胞间相互作用。因此，我将重组Shh和工具包基因中的Wnt基因这些牙源性形态发生相关的基因引入间充质细胞，并进行了类似的实验，分析了Shh和Wnt这些信号对硬质母细胞的影响。我继续分析到2011年，当使用引入重组Shh和Wnt基因的间充质细胞作为饲养细胞时，成釉细胞上皮集落计数明显增加，最大集落直径显著增大。此外，将成釉细胞接种于丝裂霉素c处理过的间充质细胞上，培养2周后成釉细胞形成结节，结节明显变大，Shh和Wnt信号组调控细胞分化的Notch1基因表达受到抑制。Wnt信号完全抑制成釉细胞Runx2/Cbfa1基因表达。今年，我制作了胶原支架、间充质细胞和成釉细胞的混合物，并将其皮下移植到裸鼠背部。我没有达到牙齿再生，因为移植混合物的致癌变化。

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.