口腔医学研究

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (7): 649-652.

• 基础研究论著 • 上一篇    下一篇

bFGF重组慢病毒对羊骨髓间充质干细胞成骨功能影响的研究

韩祥祯1,魏薇2,何惠宇1*,胡杨1,王欢欢1,邬薇薇1   

  1. 1. 新疆医科大学第一附属医院口腔修复科 新疆 乌鲁木齐 830054;
    2. 新疆医科大学第三附属医院教学科
  • 收稿日期:2014-12-15 出版日期:2015-07-28 发布日期:2015-07-28
  • 通讯作者: 何惠宇,E-mail:hehuiyu01@126.com
  • 作者简介:韩祥祯(1987~ ),男,河南人,硕士在读,住院医师,主要从事口腔修复学临床及基础的研究工作。
  • 基金资助:
    新疆维吾尔自治区科技厅科技支疆项目(目编号:201291173)

Effect of bFGF Recombinant Lentiviral Vector on the Osteogenic Potential of Goat Bone Marrow-derived Mesenchymal Stem Cells

HAN Xiang-zhen, WEI Wei, HE Hui-yu, et al   

  1. Department of Prosthodontics, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054
  • Received:2014-12-15 Online:2015-07-28 Published:2015-07-28

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摘要： 目的:探讨碱性成纤维生长因子(basic Fibroblast Growth Factor,bFGF)基因转染对体外培养的骨髓间充质干细胞(Bone Marrow mesenchymal Stem Cells, BMSCs)的增殖和骨向分化的影响,为构建组织工程骨的种子细胞做理论依据。方法:构建携带人bFGF基因的慢病毒载体,转染诱导后羊的BMSCs,得到bFGF转染组,选择未转染的BMSCs为对照组。两组细胞采用实时定量PCR技术和蛋白质印迹法技术检测Collagen-Ⅰ、OC、OPN等成骨相关基因的mRNA水平及蛋白水平相对表达量的变化情况。结果:bFGF组OPN和Collagen-Ⅰ基因mRNA水平表达量较对照组增高,差异有统计学意义(P<0.05),两组细胞在OC基因mRNA水平表达量上差异无统计学意义;bFGF组细胞在OPN、OC和Collagen-Ⅰ基因蛋白水平上表达量均高于对照组,且差异有统计学意义。结论:bFGF基因转染能增强羊BMSCs增殖和骨向分化的能力,可作为组织工程骨的种子细胞。

关键词: 碱性成纤维因子, 慢病毒载体, 种子细胞, 实时荧光定量PCR, 蛋白质印迹法

Abstract: Objective: To study the effects of bFGF transfection on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. Methods: Lentiviral vectors carrying bFGF were constructed to transfect sheep bone marrow mesenchymal stem cells. Cells were divided into two groups: bFGF group, and control group(non-transfected goat bone marrow mesenchymal stem cells). Real-time quantitative PCR and Western-blot were used to detect the changes of mRNA levels and protein levels of collagen-I, osteocalcin and osteopontin. Results: Expressions of collagen-I and osteopontin in mRNA levels were higher in transfection group than those in the control group, and the difference was statistically significant (P<0.05), But the expressions of osteocalcin in mRNA levels were not significant different in two group. Expression of OPN, Collagen- I, and OC of transfection group was higher than that in the control group, and the difference was statistically significant. Conclusion: bFGF transfection can enhance the ability of bone marrow mesenchymal stem cells on proliferation and osteogenic differentiation ,and they are suitable for seed cells for bone tissue engineering.

Key words: bFGF, Lentiviral vectors, Seed cells, Real-time, PCR, Western-blot

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