体外诱导C2C12细胞向成骨细胞的分化

Induced differentiation of C2C12 to osteoblast via adenovirus-mediated Cbfa1 in vitro

目的成骨细胞特异性转录因子a1(core binding factor a1,Cbfa1)通过调节生长因子和骨特异性细胞外基质蛋白的基因表达而参与成骨细胞的分化和骨发育过程。文中构建成Cbfa1,以腺病毒载体转染成肌细胞C2C12,为种子细胞构建组织工程化骨。方法体外培养小鼠成肌细胞C2C12,用重组腺病毒质粒pAd-IL-31介导Cbfa1/Osf2基因瞬时转染小鼠成肌C2C12细胞,Western blot检测Cbfa1蛋白表达。结果 Cbfa1蛋白表达、碱性磷酸酶(alkaline phosphatase,ALP)活性测定、骨钙素(osteocalcin,OCN)分泌量以及茜素红染色感染组明显高于对照组。结论成肌细胞C2C12可以作为种子细胞构建组织工程化骨。

Objective Osteoblast core binding factor al （ Cbfal ） plays a role in osteoblast differentiation and development by regulating the gene of growth factor and extracellular matrix proteins. Recombinant adenovirus vector mediated Cbfal was transferred to myohlast C2C12 to construct the tissue-engineered bone. Methods The myoblast C2C12 was cultured in vitro, and then transiently transfected with recombinant adenovirus vector pAd-IL-31 mediated-Cbfal/Osf2. Western blot was used to detect the expression of Cbfal. Results Compared with the control group, the expression of Cbfal, activity of alkaline phosphtase （ALP） , secretory volume of osteocalcin （OCN） and staining via alizarin bordeaux were higher in the transfection group. Conclusion Myoblast C2C12 acts as a seed cell for constructing tissue-engineered bone.