Cloning, expression and characterization of gene encoding human stem cell growth factor-alpha and its synergetic effect with rhGM-CSF on proliferation of human umbilical cord mesenchymal stem cells.
- Author:
Xinlei PENG
1
;
Yanyan MA
;
Jing RONG
;
Zhenling ZHAO
;
Bo HAN
;
Wei CHEN
;
Yangfei XIANG
;
Qiuying LIU
;
Yifei WANG
;
Zhe REN
;
Xiangrong ZHOU
;
Haijia CHEN
Author Information
1. Guangdong Provincial Key Laboratory of Bioengineering Medicine, Biomedicine Research and Development Centre of Jinan University, Guangzhou 510632, China.
- Publication Type:Journal Article
- MeSH:
Cell Proliferation;
drug effects;
Cells, Cultured;
Cloning, Molecular;
Drug Synergism;
Escherichia coli;
genetics;
metabolism;
Granulocyte-Macrophage Colony-Stimulating Factor;
pharmacology;
Humans;
Mesenchymal Stromal Cells;
cytology;
metabolism;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
pharmacology;
Stem Cell Factor;
biosynthesis;
genetics;
Umbilical Cord;
cytology
- From:
Chinese Journal of Biotechnology
2011;27(11):1667-1676
- CountryChina
- Language:Chinese
-
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.
