端粒延伸替代途径在人骨髓间充质干细胞中的表达和作用  

Expression of alternative lengthening telomere in human bone marrow mesenchymal stem cells

作　　者：赵强[1] 张丽红[2] 吴珊[2] 何旭[2] 石英爱[2] 

机构地区：[1]吉林大学第一医院儿外科,吉林长春130021 [2]吉林大学基础医学院病理生物学教育部重点实验室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2010年第1期27-29,I0001,共4页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30700872);吉林省科技厅科技发展计划项目资助课题(200705369)

摘　　要：目的:分离、培养人骨髓间充质干细胞(hMSCs),观察端粒延伸替代途径相关的白血病前髓淋巴细胞小体(PML)在骨髓间充质干细胞中的表达情况,明确其在hMSCs自我更新和分裂增殖过程中的作用。方法:Percoll梯度离心法分离培养hMSCs,免疫细胞化学染色方法检测PML小体在hMSCs中的表达率。结果:P1、P3和P5代hMSCs中未见明显PML小体表达;P7代hMSCs中可见PML小体在细胞胞核中呈阳性表达,表现为胞核内出现棕黄色颗粒,表达率为(16.1±0.6)%;P9和P11代hMSCs中亦可见PML小体表达,阳性率分别为(16.8±2.6)%和(45.8±9.5)%,P11代hMSCsPML小体表达率高于P7代hMSCs(P<0.05)。结论:随着hMSCs传代次数的增加,端粒延伸替代途径逐渐出现发挥其延长端粒长度的作用,进而维持细胞的增殖和自我更新的能力。Objective To isolate and cultivate human bone marrow mesenchymal stern cells （hMSCs） in vitro, and observe the expression of alternative lengthening telomere related promyelocytic leukemia protein （PML） in hMSCs, in order to evaluate its role in self-renewal and division growth processes of bMSCs. Methods hMSCs were isolated from bone marrow by Percoll density gradient centrifugation. Immunocytochemistry staining was used to detect the expression rate of PML body in hMSCs. Results There were no visible expressions of PML body in passage 1, 3 and 5 of cultured hMSCs. There was positive expression of PML body in passage 7 hMSCs, there were brown particles in nuclei of hMSCs, the positive rate was （16. 1± 0.6）%. There were also positive expressions of PML body in passage 9 and 11 hMSCs, the positive rates were （16.8±2.6）% and （45.8±9.5）%, respectively; there were significant differences between passage 9, 11 and passage 7 hMSCs （P〈0.05）. Conclusion Following long- time culture of hMSCs in vitro, ALT will appear in hMSCs to promote the lengthening of telomere, and maintain the ability of proliferation and self-renewal of hMSCs.

关 键 词：骨髓 间质干细胞 端粒酶 免疫组织化学 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]