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  • 汤立旦1,余建2,王冲2,来晓瑜2.人骨髓间质干细胞向脂肪细胞诱导分化过程中端粒酶活性的变化[J].第二军医大学学报,2011,32(2):123-127    [点击复制]
  • TANG Li-dan1, YU Jian2, WANG Chong2, LAI Xiao-yu2.Changes of telomerase activity during differentiation of human mesenchymal stem cells into adipocytes[J].Acad J Sec Mil Med Univ,2011,32(2):123-127   [点击复制]
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人骨髓间质干细胞向脂肪细胞诱导分化过程中端粒酶活性的变化
汤立旦1,余建2,王冲2,来晓瑜2
0
(1.浙江医学高等专科学校医学二系,杭州 310053; 2.浙江大学医学院附属第一医院血液科,杭州 310003)
摘要:
目的观察体外培养人骨髓间质干细胞(mesenchymal stem cells, MSCs)成脂分化过程中端粒酶活性的变化规律,并探讨其可能的机制。方法通过贴壁培养法从人骨髓中分离MSCs,利用造血干细胞相关表面抗体对MSCs进行表型鉴定,利用相应诱导分化剂,诱导MSCs向成骨细胞、脂肪细胞和神经元样细胞分化,分析其多向分化潜能。以端粒重复序列扩增法(TRAP)检测MSCs及MSCs诱导成脂分化过程中端粒酶活性;以Western印迹法检测MSCs及MSCs诱导成脂分化过程中端粒酶反转录酶(hTERT)及端粒相关蛋白\[端粒重复序列结合因子1(TRF1)、端锚聚合酶1(Tankyrase 1)\]的表达水平。结果TRAP法检测传代培养的MSCs端粒酶呈阴性表达,而MSCs诱导成脂分化过程中,端粒酶活性明显升高,以第7天最高(P<0.05),其后呈下降趋势;Western印迹法检测显示,hTERT在MSCs中有微弱表达,在MSCs诱导成脂分化过程中,hTERT和Tankyrase 1表达水平升高,以第7天最高,此后逐渐下降,而TRF1表达水平保持不变。结论体外培养的MSCs端粒酶呈微弱表达,MSCs在成脂诱导分化过程中,端粒酶活性先升高,其后逐渐下降,而Tankyrase 1可能在其中发挥重要作用。
关键词:  间质干细胞  细胞分化  脂肪细胞  端粒酶
DOI:10.3724/SP.J.1008.2011.0123
投稿时间:2010-11-15修订日期:2011-01-12
基金项目:
Changes of telomerase activity during differentiation of human mesenchymal stem cells into adipocytes
TANG Li-dan1, YU Jian2, WANG Chong2, LAI Xiao-yu2
(1. Department of Medicine Ⅱ, Zhejiang Medical College, Hangzhou 310053, Zhejiang, China; 2. Department of Hematology, the First Affiliated Hospital, Zhejiang University Medical School, Hangzhou 310003, Zhejiang, China)
Abstract:
\[Abstract\]ObjectiveTo study the telomerase activity changes during the differentiation of human mesenchymal stem cells (MSCs) into adipocytes and the related mechanism. MethodsHuman MSCs were isolated from human bone marrow by plastic adhesion and were characterized by immune phenotyping. The multilineage differentiation potential of MSCs was confirmed by inducing MSCs to differentiate into osteoblasts and neuron-like cells by corresponding agents. Telomeric repeat amplification protocol (TRAP) assay was performed to examine telomerase activity in MSCs during adipogenic differentiation of MSCs; Western blotting analysis was used to examine the protein expression levels of hTERT, TRF1, and Tankyrase 1. ResultsTRAP showed that telomerase was negative in MSCs, but greatly up-regulated during the adipogenic differentiation process, peaked on the 7th day after adipogenic differentiation of MSCs(P<0.05), and then decreased thereafter. Western blotting analysis showed slight hTERT expression in MSCs, and the expression of hTERT and Tankyrase 1 increased during the adipogenic differentiation process, peaked on the 7th day, and gradually decreased thereafter. The expression of TRF1 remained unchanged. ConclusionTelomerase is weakly expressed in MSCs; the expression is increased during the adipogenic differentiation process of MSCs and then gradually decreases; and Tankyrase 1 may play an important role during the process.
Key words:  mesenchymal stem cells  cell differentiation  adipocytes  telomerase