The clone of OPG and its effects on the differentiation of bome morrow stem cells

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The clone of OPG and its effects on the differentiation of bome morrow stem cells

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- The clone of OPG and its effects on the differentiation of bome morrow stem cells
- Journal of Chongqing Medical University Vol. 34, Issue 5, Pages: 532-536(2009)
- 作者机构：
  
  重庆医科大学附属第二医院骨科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R346
- Published：2009，
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[1]陈富,徐希彦,柯珍勇,尹良军,郭常辉,王雪燕,林春阳,邓忠良.骨保护素的克隆及对骨髓间充质干细胞分化的影响[J].重庆医科大学学报,2009,34(05):532-536.

CHEN Fu, et al. The clone of OPG and its effects on the differentiation of bome morrow stem cells. [J]. Journal of Chongqing Medical University 34(5):532-536(2009)
[1]陈富,徐希彦,柯珍勇,尹良军,郭常辉,王雪燕,林春阳,邓忠良.骨保护素的克隆及对骨髓间充质干细胞分化的影响[J].重庆医科大学学报,2009,34(05):532-536. DOI：

CHEN Fu, et al. The clone of OPG and its effects on the differentiation of bome morrow stem cells. [J]. Journal of Chongqing Medical University 34(5):532-536(2009) DOI：

摘要

目的:克隆人骨保护素（Osteoprotegerin

OPG）基因并构建其重组腺病毒

观察其对BMSCs分化的影响.方法:RT-PCR方法从人肝克隆OPG基因全长cDNA

利用穿梭质粒将OPG基因重组获得复制缺陷型重组腺病毒AdOPG;分离培养兔骨髓间充质干细胞

使用AdOPG感染兔BMSCs

以RT-PCR及Western Blot的方法鉴定BMSCs对OPG的表达。比色测定和细胞碱性磷酸酶染色法观察AdOPG感染BMSCs后5d时碱性磷酸酶（ALP）的表达。结果:克隆获得人OPG基因

测序与Gene bank一致

构建的重组腺病毒AdOPG滴度达109efu/ml

AdOPG感染BMSCs5d后ALP活性值为（21024±507）IU

AdGFP对照组为（3079±89）IU

空白对照组为（2156±78）IU。结论:骨保护素有利于骨髓间充质干细胞向成骨细胞方向分化。

Abstract

Objective:Toclone the OPGgene ofhuman beingand construct its recombinant adenovirus and then observe its effects on the differentiation of bone marrow stem cells(BMSCs).Methods:The cDNAs of human OPG in hepatic tissue were obtained by using RT-PCR method.The cDNAs then were recombined with competent homologous cells to form the AdOPG and the shuttle vector was used during this stage.The rabbit BMSCs were isolated and cultured in vitro

and were infected with AdOPG.The OPG expression of BMSCs was identified by RT-PCR and Western blot.The expression of ALP was detected by chromatometry and ALP dyeing five days after the infection.Results:The OPG gene of human being was successfully cloned and its sequence consistent with the Gene bank.The titre of constructed AdOPG can reach 109 efu/ml.The ALP activity of BMSCs was(21 024±507)IU five days after the infection of AdOPG

whereas the value was(3 079±89)IU in the control group and(2 156±78)IU in the blank control group.Conclusion:The OPG can induce the osetogeinc differentiation of BMSCs.

关键词

腺病毒人骨保护素骨髓间充质干细胞

Keywords

adenovirusHuman OPGBone marrow stem cells

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