端粒酶活性在骨髓间充质干细胞中的表达

目的：探讨骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells MSCs)端粒酶活性的表达并检测其部分生物学特性。方法：取正常成人骨髓用含10%新生牛血清的LG-DMEM培养液培养、扩增。流式细胞仪行细胞表面抗原检测，细胞化学染色鉴定其生物学特性，观察其在地塞米松、β-甘油磷酸钠、维生素C特定培养条件下向成骨细胞分化的能力，采用TRAP（Telomerase repeat amplification protoco1 assay)-银染法测定其端粒酶的活性。结果：分离培养获得的贴壁细胞，碱性磷酸酶染色阳性。流式细胞仪检测CD34、CD45呈阴性表达，CD29、CD44呈阳性表达。经向成骨细胞诱导3周后，可得到成骨细胞，经茜素红染色得到证实。TRAP-银染法证实MSCs表达一定的端粒酶活性。结论：体外分离培养的MSCs可以向成骨分化，表达一定的端粒酶活性，具有干细胞的特性。

Telomerase activation of bone marrow mesenchymal stem cells

Objective： To explore the elomerase activation of bone marrow-derived mesenchymal stem cells(MSCs) and its biology characteristics. Methods: Bone-derived MSCs were cultured in Dulbecco′s modified eagle′s medium(DMEM) with low glucose containing 10 % new bovine serum. Flow cytometer(FCM) was used to determine the expression of cell surface molecules, and cytochemical staining was used to identify their biology characteristics. Osteogenic differentiation was assessed in low glucose DMEM containing dexamethasone, β-sodium glycerophosphate and vitamin C. Telomerase repeat amplification protocol assay（TRAP） argentation method was used to determine the telomerase activation. Results: MSCs could be isolated and cultured from bone marrow and they were negative for CD34 and CD45 but positive for CD29 and CD44. Three weeks after inducement by osteoblast, they were positive for alizarin red staining and expressed telomerase activation. Conclusion： MSCs isolated and cultured from adult human bone marrow can be induced into osteoblasts, and express telomerase activation and have stem cell biology characteristics.