不同直径牙胚细胞增殖和骨向分化能力的评估

Proliferation and osteogenic differentiation of tooth germ cells with different diameters

目的:比较大鼠不同直径牙胚细胞增殖和骨向分化能力的差异。方法:酶消化法分离、培养SD大鼠牙胚细胞,经13μm标准细胞筛分选后获得大、小2种直径的细胞;并分别采用MTT法检测2种细胞的增殖能力;碱性磷酸酶(ALP)试剂盒、茜素红染色法及qRT-PCR检测两者的骨向分化能力。结果:小细胞的增殖能力较低(P<0.05);培养3、7 d后,小细胞组的ALP活性均明显高于大细胞组(P<0.05);培养14 d后,小细胞组钙化结节数量明显高于大细胞组;qRT-PCR检测结果显示,小细胞组中Ocn、Osx、Runx2各矿化相关基因的表达水平均明显高于大细胞组(P<0.05)。结论:牙胚细胞中直径小于13μm的细胞比直径大于13μm的细胞增殖活性低,分化能力强。

AIM： To compare the proliferation and osteogenic differentiation of rat tooth germ cells （ rT-GCs） with different diameters.METHODS：rTGCs were isolated enzymatically from Sprague-Dawley rats and divided into larger and smaller populations using a 13 μm cell sieve.MTT assay was performed to evaluate the cell prolifera-tion.Alkaline phosphotase （ ALP） kits, alizarin red staining and quantitative PCR were used to investigate the osteo-genic differentiation capacity of the cells.RESULTS： The larger rTGCs（ rTGCs-Large） showed greater proliferation rate than the smaller counterparts （ rTGCs-Small） .However, rTGCs-Small group presented higher ALP activity at 3 and 7 d after culture and more calcified nodules than rTGCs-Large group at 14 d after culture.Moreover, rTGCs-Small group expressed stronger osteogenic markers Ocn, Osx and Runx2 than rTGCs-Large group.CONCLUSION：rTGCs-Small have lower proliferation capacity but stronger osteogenic differentiation potential than rTGCs-Large group.