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A constructing method of a bone marrow stem cell expressing an osteogenic gene

A bone marrow mesenchymal and osteogenic gene technology, applied in genetic engineering, plant gene improvement, botanical equipment and methods, etc., to achieve the effects of good cell growth, simple operation, and improved expression level

Inactive Publication Date: 2015-08-12
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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Problems solved by technology

Currently, there is no method to infect bone marrow mesenchymal stem cells expressing osteogenic genes using hypoxia-inducible factor 1α recombinant lentivirus

Method used

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  • A constructing method of a bone marrow stem cell expressing an osteogenic gene
  • A constructing method of a bone marrow stem cell expressing an osteogenic gene
  • A constructing method of a bone marrow stem cell expressing an osteogenic gene

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Embodiment Construction

[0019] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0020] The lentiviral vector is obtained by transformation of human immunodeficiency virus (humanimmunodeficiency virus, HIV). The lentiviral vector system includes expression vector and packaging plasmid. Integrate the required genetic information. On the basis of HIV, the four auxiliary genes vif, vpr, vpu and nef were deleted, and at the same time, a plasmid expressing vesicular stomatitis virus (VSV) glycoprotein G and amphotropic murine leukemia virus (MLV) envelope protein Env were used The plasmids that express the HIV-1 envelope protein Env are replaced re...

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Abstract

The invention is suitable for the field of biological medicines, and provides a constructing method of a bone marrow stem cell expressing an osteogenic gene. The method utilizes a lentivirus carrying a hypoxia-inducible factor 1alpha to infect a bone marrow stem cell and to express the osteogenic gene. The method includes following steps: constructing a lentivirus expression vector Lenti-HIF-1alpha-eGFP; packaging the lentivirus expression vector Lenti-HIF-1alpha-eGFP carrying the hypoxia-inducible factor 1alpha; and infecting the bone marrow stem cell with the lentivirus expression vector Lenti-HIF-1alpha-eGFP to obtain the infected bone marrow stem cell capable of expressing the osteogenic gene.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a construction method for bone marrow mesenchymal stem cells expressing osteogenic genes. Background technique [0002] Large-scale bone defects caused by trauma, tumors, inflammation, etc. are long-term clinical problems and seriously endanger human health. Traditional treatment methods such as autologous bone transplantation have problems such as lack of bone mass, allogeneic bone grafts may spread diseases, and xenogeneic allogeneic bone grafts will cause immune rejection. Therefore, bone tissue engineering has become a new strategy for the treatment of large-scale bone defects. . At present, in animal experiments, when tissue engineered bone repairs small-scale bone defects, the fracture healing effect is very good, but when repairing large-scale bone defects, avascular necrosis often occurs in the central part. Therefore, it is very difficult to simply use tissue eng...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867C12Q1/68
Inventor 熊建义王大平张巨峰付志杰李庆庆廖福朋
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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