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Chinese Journal of Biotechnology ; (12): 1667-1676, 2011.
Article in Chinese | WPRIM | ID: wpr-304533

ABSTRACT

To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.


Subject(s)
Humans , Cell Proliferation , Cells, Cultured , Cloning, Molecular , Drug Synergism , Escherichia coli , Genetics , Metabolism , Granulocyte-Macrophage Colony-Stimulating Factor , Pharmacology , Mesenchymal Stem Cells , Cell Biology , Metabolism , Recombinant Fusion Proteins , Genetics , Pharmacology , Stem Cell Factor , Genetics , Umbilical Cord , Cell Biology
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