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[The stable expression of human tissue-type plasminogen activator gene mediated by lipofectamine in human vein endothelial cell line cells].
- Source :
-
Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering = Shengwu yixue gongchengxue zazhi [Sheng Wu Yi Xue Gong Cheng Xue Za Zhi] 2007 Dec; Vol. 24 (6), pp. 1330-3. - Publication Year :
- 2007
-
Abstract
- We have established a human umbilical vein endothelial cell (HUVEC) line monoclonal cells with the stable expression of human tissue-type plasminogen activator (t-PA) gene to provide a basis for further study on the vascular tissue engineering. Recombinant plasmid pcDNA3. 1-Myc-His B (-)/t-PA was constructed by insertion of t-PAcDNA originated from PBS/t-PA into eukaryotic expression vector pcDNA3. 1-Myc-His B(-) and transfected into hUVEC line cells mediated by lipofectamine. The positive clones were obtained by the screen of G418. The transcription and expression of t-PA gene were investigated by RT-PCR and Western blotting respectively. The t-PA activity was measured by chromogenic substrate assay. The positive clone cells which transcripted the mRNA of t-PA gene was obtained by RT-PCR. Immunoreactive human t-PA of the medium was significantly increased in the group of transfected gene when compared with that in the controlled and transfected plasmid without t-PA gene group. The biological activity of the protein of the t-PA in the media was increased significantly in the positive clone cells with t-PA gene transfected. The HUVEC line monoclonal cells with the stable expression of t-PA gene was established successfully.
- Subjects :
- Cell Line
Humans
RNA, Messenger biosynthesis
RNA, Messenger genetics
Recombinant Proteins biosynthesis
Recombinant Proteins genetics
Tissue Engineering
Tissue Plasminogen Activator genetics
Transfection
Human Umbilical Vein Endothelial Cells metabolism
Lipids pharmacology
Tissue Plasminogen Activator biosynthesis
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1001-5515
- Volume :
- 24
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering = Shengwu yixue gongchengxue zazhi
- Publication Type :
- Academic Journal
- Accession number :
- 18232487