人脱落乳牙牙髓干细胞促进破骨细胞形成的体外实验研究

The study of stem cells from human exfoliated deciduous teeth promote osteoclastogenesis in vitro

目的:通过间接共培养的方法初步探讨人脱落乳牙牙髓干细胞(stem cells from human exfoliated deciduous teeth,SHED)在体外对破骨前体细胞分化的影响。方法:建立SHED与破骨前体细胞(外周血单个核细胞,peripheral blood mononuclear cells,PBMCs)的间接共培养模型,利用TRAP染色观察PBMCs向破骨细胞样细胞分化后的细胞数目与形态,利用qPCR技术和Western Blot技术检测破骨相关基因TRAP、CTSK、c-Fos、TRAF6的表达,以评估PBMCs向破骨细胞样细胞分化的情况。结果:与对照组相比,SHED与PBMCs间接共培养模型中,TRAP染色阳性、具有多个核的破骨细胞样细胞数量显著增多,破骨相关基因TRAP、CTSK、c-Fos、TRAF6在PBMCs中的表达水平显著上调。结论:SHED能够以细胞非接触的方式促进破骨前体细胞向破骨细胞样细胞分化。

Objective： To investigate the effect of stem cells from human exfoliated deciduous teeth（ SHED） on differentiation of osteoclast precursors in vitro by using indirect co-culture approach. Methods： The indirect co-culture system of SHED and osteoclast precursor cell line peripheral blood mononuclear cells（ PBMCs） was established. In order to evaluate the osteoclastic differentiation of PBMCs after co-culture,the morphology of the osteoclast-like cells was observed by TRAP staining and the number of TRAP＋cells was counted. The expression of TRAP,CTSK,c-Fos,TRAF6 in PBMCs was evaluated by q PCR and Western Blot. Results： SHED promoted the differentiation of PBMCs into TRAP＋osteoclast-like cells and the expression of TRAP,CTSK,c-Fos,TRAF6 in PBMCs,compared with control group. Conclusion： SHED can promote the differentiation of osteoclast precursors into osteoclast-like cells by noncontact co-culture.