《中国康复理论与实践》 ›› 2005, Vol. 11 ›› Issue (01): 1-3.

• 专题 •    下一篇

阳离子脂质体介导重组人骨形态发生蛋白-2基因转染的兔骨骼肌干细胞体外成骨分化的实验研究

尚咏1; 伍骥1; 卢世璧2; 袁玫2   

  1. 1.空军总医院骨科 北京市,100036;2.解放军总医院骨科 北京市,100853
  • 收稿日期:2004-12-01 出版日期:2005-01-25 发布日期:2005-01-25

Osteoinductivity of recombinent human bone morphogenetic protein-2 gene mediated with cationic liposome transfer in skeletal muscle stem cells of rabbits in vitro

PEI Hai-tao,SUN Jin-ping   

  1. Emergency Neurology Department of the Hospital Affiliat- ed to Medical College of Qingdao University, Qingdao266003, Shandong, China
  • Received:2004-12-01 Published:2005-01-25 Online:2005-01-25

摘要: 目的初步探讨阳离子脂质体介导的重组人骨形态发生蛋白-2(rhBMP-2)基因对兔骨骼肌干细胞(SMSCs)体外成骨活性的影响。方法PEGFP-rhBMP-2的扩增、浓度及纯度鉴定。脂质体包裹质粒DNA,转染SMSCs后测定转染率。检测rhBMP-2、碱性磷酸酶(ALP)和Ⅰ型胶原的表达。结果脂质体介导 PEGFP-rhBMP-2 SMSCs转染后最大转染率为14.18%,rhBMP-2、ALP和Ⅰ型胶原的表达呈阳性,而PEGFP转染的细胞和未染细胞rhBMP-2、ALP和Ⅰ型胶原的表达弱阳性。结论SMSCs是一种较理想的骨组织工程种子细胞。脂质体介导质粒rhBMP-2的基因转移安全性好,但转染率相对较低。

关键词: 骨形态发生蛋白, 阳离子脂质体, 骨骼肌干细胞, 基因治疗, 骨缺损,

Abstract: ObjectiveTo evaluate the osteoinductivity of cationic liposome mediated recombined human bone morphogenetic protein-2 gene (rhBMP-2) transfer in skeletal muscle stem cells (SMSCs) of rabbits. MethodsSMSCs were transfered by pAGFP rhBMP-2 liposome complex. Gene transferred SMSCs were evaluated with immunohistochemistry.ResultsThe transfection rate of SMSCs transferred by cationic liposome mediated pAGFP-rhBMP-2 was 14.18%. Positive stain of rhBMP-2, alkaline phosphatase(ALP),collegenⅠhad been found in the transferred SMSCs.ConclusionQuantity of SMSCs is enough to be target cells of gene therapy. Cationic liposome can mediated bone morphogenetic protein-2 gene transfected SMSCs in vitro. Compared with AdrhBMP-2,the transfection rate of liposome mediated bone morphogenetic protein-2 gene transfected SMSCs is lower.

Key words: bone morphogenetic protein, cationic liposome, skeletal muscle stem cells, gene therapy, bone defect, rabbit