目的　通过重建端粒酶活性延长人成纤维细胞寿命 ,并对其成骨潜能进行研究 ,为解决组织工程骨修复种子细胞老化问题提供实验依据。方法　将人端粒酶催化亚基 (h TERT)基因用电穿孔法导入正常人原代成纤维细胞 ,用 TRAP- PCR检测细胞端粒酶活性 ,用β-半乳糖苷酶活性测定评价细胞衰老情况。在此基础上用骨形成蛋白(BMP- 2 )和肿瘤坏死因子 -α(TNF-α)联合诱导已重建端粒酶活性的成纤维细胞在体外培养条件下成骨 ,用四环素活体标记和茜素红染色显示钙盐结节形成。结果　转染 h TERT的人成纤维细胞能稳定表达端粒酶活性 ,培养超过 5 0代后细胞仍保持β-半乳糖苷酶阴性。经 BMP- 2和 TNF-α诱导后 ,转染后传 80代的人成纤维细胞仍可形成四环素标记和茜素红染色均为阳性的钙盐结节。结论　重建端粒酶活性、寿命延长的人成纤维细胞仍然维持了其本身所具有的成骨潜能 OBJECTIVE: To prolong the lifespan of human fibroblasts by reconstituting telomerase activity and to study their osteogenic potential to provide experimental basis for solving the problem of tissue engineering bone repair of seed cells aging. Methods The human telomerase catalytic subunit (h TERT) gene was introduced into normal human primary fibroblasts by electroporation. The telomerase activity was detected by TRAP-PCR and cell senescence was evaluated by β-galactosidase activity assay Happening. On this basis, fibroblasts that have been induced to reconstitute telomerase activity by osteogenic protein (BMP-2) and tumor necrosis factor-α (TNF-α) are osteoblasts cultured in vitro, labeled with tetracycline and labeled with Su-red staining showed calcium nodule formation. Results Human fibroblasts transfected with h TERT could stably express telomerase activity. When cultured for more than 50 generations, the cells remained negative for β-galactosidase. After induced by BMP-2 and TNF-α, 80-day-old human fibroblasts can still form tetracycline-labeled and alizarin red-stained calcium nodules. Conclusion The human fibroblasts with telomerase activity and prolonged lifespan maintained their osteogenic potential