再生障碍性贫血患者骨髓间充质干细胞生物学特性的初步研究

目的:研究再生障碍性贫血(aplasticanemia,从)患者骨髓间充质干细胞(mesenchymalstemcells,MSCs)的生物学特性和初步探讨其异常和AA发生的可能关系。方法:取AA患者骨髓间充质干细胞,测定其生长曲线和倍增时间;流式细胞仪检测其细胞周期和免疫表型;体外定向诱导其向脂肪、成骨、内皮和神经细胞分化;用real-timePCR及油红O染色法比较AA和正常对照组MSCs的成脂分化的不同。结果:AA患者和正常成人的MSCs均呈梭形贴壁生长;AA组细胞倍增时间长于对照组;CD105、CD44、CD29、CD106、FlK-1均阳性;96．51％的细胞处在G0／G1期;AA患者的MSCs保持了多向分化潜能,体外诱导形成脂滴较对照组早,诱导早期的脂蛋白脂酶表达增高。结论:再生障碍性贫血患者的骨髓间充质干细胞增殖能力较正常成人弱,骨髓间充质干细胞的易成脂性可能参与了再障的发病环节。

Primary Study of the Characteristics of Mesenchymal Stem Cells Derived from Bone Marrow of Aplastic Anemia Patients

Objective: To study the characterristics of mesenchymal stem cells (MSCs) from bone marrow of aplastic anemia (AA) patients and explore the relation between the abnormity of MSCs and the pathological mechanisms of aplastic anemia. Methods: MSCs from bone marrow of AA patients and healthy adults (normal control) were isolated. The growth curve and cell-doubling time were measured.The immunophenotype and cell cycle were assessed by flow cytometry. The multilineage differentiation capacities were investigated. The differentiation potential into adipocytes was evaluated by staining of oil red O and real-time PCR. Results: MSCs from bone marrow of aplastic anemia (AA) .patients and normal control were fibroblast-like cells. The cell-doubling time of MSCs from AA group was longer than that from control group. Flow cytometry analysis showed these cells were CD105, CD44, CD29, CD106 and Flk-1 possitive. Cell cycle analysis revealed that more than 90% of cells were in the G0/G1 phases. Under the appropriate culture conditions, MSCs from AA patient could differentiate into osteogenic,adipogenic, endothelial and neural lineages. Compared with normal control, MSCs from AA patients could differentiate more easily into adipocytes. Conclusion: The growth capacity of MSCs from AA patients is lower than that from control group. The increased adipogenic capacitiy of MSCs may contribute to the pathogenesis of AA.