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Human bone morphogenetic protein-2 gene transfer induces human mesenchymal stem cell proliferation and differentiation in vitro

机译:人骨形态发生蛋白2基因转移在体外诱导人间充质干细胞增殖和分化

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摘要

Objective: To identify eukaryotic expression vector of human bone morphogenetic protein 2 pcDNA3/BMP2, verify its expression in transfected human mesenchymal stem cells (hMSCs) and the effect on hMSCs differentiation.Methods: The BMP2 gene was cloned into a eukaryotic expression vector pcDNA3. Transfected the recombinant into hMSCs by liposome. Immunnohistochemistry and in situ hybridization methods were used to identify the expression of BMP2 mRNA and protein; ALP and Von Kossa stains were performed to identify the BMP2 gene differentiated effect on the hMSCs. Results: The pcDNA3/BMP2 fragments were as large as theory. BMP2 mRNA and protein were expressed and synthesized both in 48 h and 4 weeks after transfection, the ALP and Ca deposit exhibition, which marked the osteogenic lineage of hMSCs,were enhanced and sped. Conclusion: Transfection of pcDNA3/BMP2 is able to provide transient and persistent expression in hMSCs, and promote the MSCs differentiation to osteogenic lineage.
机译:目的:鉴定人骨形态发生蛋白2 pcDNA3 / BMP2的真核表达载体,验证其在转染的人间充质干细胞(hMSCs)中的表达及其对hMSCs分化的影响。方法:将BMP2基因克隆到真核表达载体pcDNA3中。通过脂质体将重组体转染到hMSC中。采用免疫组织化学和原位杂交方法鉴定BMP2 mRNA和蛋白的表达。进行ALP和Von Kossa染色以鉴定BMP2基因对hMSC的分化作用。结果:pcDNA3 / BMP2片段与理论值一样大。转染后48 h和4周,BMP2 mRNA和蛋白表达并合成,ALP和Ca沉积物的出现,标志着hMSCs的成骨谱系得到增强和加速。结论:pcDNA3 / BMP2转染能够在hMSCs中提供瞬时和持续表达,并促进MSCs向成骨细胞系的分化。

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  • 作者单位

    Department of Orthopaedic Surgery, Tangdu Hospital, Forth Military Medical University, Xi' an 710038, China;

    Department of Orthopaedic Surgery, Tangdu Hospital, Forth Military Medical University, Xi' an 710038, China;

    Department of Orthopaedic Surgery, Tangdu Hospital, Forth Military Medical University, Xi' an 710038, China;

    Department of Orthopaedic Surgery, Tangdu Hospital, Forth Military Medical University, Xi' an 710038, China;

    Department of Orthopaedic Surgery, Tangdu Hospital, Forth Military Medical University, Xi' an 710038, China;

    Department of Orthopaedic Surgery, Tangdu Hospital, Forth Military Medical University, Xi' an 710038, China;

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  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 外科学;
  • 关键词

    bone morphogenetic protein(BMP2); transfection; human mesenchymal stem cells (hMSCs); osteogenic differentiation;

    机译:骨形态发生蛋白(BMP2);转染;人间充质干细胞(hMSCs);成骨分化;
  • 入库时间 2022-08-19 03:48:25
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