BrdU体内示踪骨髓基质干细胞生物学状态的效果分析

目的:探讨5-溴脱氧尿嘧啶核苷(Brd U)体内示踪骨髓基质干细胞(BMSCs)生物学状态的效果。方法:抽取健康成年比格狗骨髓,在传代培养中进行Brd U标记并鉴定,体外实验中测定细胞周期、凋亡率和细胞活力;在体内实验中将标记Brd U的骨髓基质干细胞植入自体股骨头缺损处,另一侧单纯植入自体骨作为对照,记录成骨量与分子标记物的表达情况。结果:骨髓基质干细胞的Brd U体外标记率为85.2%。Brd U组的细胞凋亡率为3.62±1.33%,未标记组为3.52±1.08%;Brd U组与未标记组的细胞成活率分别为96.31±1.39%和95.20±2.10%,两组对比差异均无统计学意义(P0.05)。移植侧Brd U标记的骨髓基质干细胞免疫组化观察可见Brd U免疫组化染色阳性,阳性率为81.6%。骨髓基质干细胞移植侧缺损区的骨钙素、Ⅰ型胶原阳性细胞表达数量与强度明显高于对照侧缺损区;骨髓基质干细胞移植侧成骨量为17.46±2.12%,对照侧为9.06±1.24%,两两对比差异有统计学意义(P0.05)。结论:Brd U在体外示踪骨髓基质干细胞能有效反映细胞的生物学状态,体内示踪显示移植的骨髓基质干细胞能成活,能促进骨组织形成和坏死骨修复。

Effect of BrdU Tracing in Vivo on the Biological Status of Bone Marrow Stromal Stem Cells

ABSTRACT Objective: To investigate the effects of 5- bromodeoxyuridine (BrdU) tracing in vivo on the biological status of bone marrow stromal stem cells(BMSCs). Methods: Healthy adult beagle dog bone marrow was extracted, then cultured and indentified by BrdU markers, the cell cycle, apoptosis rate, cell viability were determined in vitro. The stem cells were given implantation of autogenous femoral head defect by BMSCs labeled BrdU in vivo, the other side were given implantation of autogenous bone as the control, the bone mass and molecular markers expression were recorded. Results: The BrdU labeling rate of BMSCs in vitro was 85.2%. The apoptosis rate of BrdU group was 3.62±1.33%, the unlabeled group was 3.52±1.08%. The cell survival rate of BrdU group and unlabeled group were 96.31±1.39% and 95.20±2.10%(P>0.05). Immunohistochemical staining of BrdU labeled BMSCs showed positive BrdU staining, the positive rate was 81.6%. The expression quantity and the intensity of osteocalcin and collagen in the transplantation bone side were high- er than those of the control side. The bone mass in the transplantation bone side was 17.46±2.12%, which was 9.06±1.24% in the con- trol(P<0.05). Conclusion: BrdU labeled BMSCs could effectively reflect the biological state of cells in vitro, it showed that the BMSCs transplantation could survive in vivo, it could promote the bone formation and repair of necrotic bone.