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 / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1667-1676, 2011.
Article
em Zh
| WPRIM
| ID: wpr-304533
Biblioteca responsável:
WPRO
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.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Farmacologia
/
Cordão Umbilical
/
Proteínas Recombinantes de Fusão
/
Células Cultivadas
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Fator Estimulador de Colônias de Granulócitos e Macrófagos
/
Clonagem Molecular
/
Fator de Células-Tronco
/
Biologia Celular
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Proliferação de Células
/
Sinergismo Farmacológico
Limite:
Humans
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2011
Tipo de documento:
Article