Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (14): 2540-2544.doi: 10.3969/j.issn.1673-8225.2011.14.016

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Expression and secretion of hepatocyte growth factor on bone marrow mesenchymal stem cells stimulated by tumor necrosis factor alpha

Lu Zheng-feng, Jiang Hai-xing, Qin Shan-yu, Xiao Jian, Zhang Jun-hong, Meng Yun-chao   

  1. Department of Gastroenterology Medicine, the First Affiliated Hospital of Guangxi Medical University, Nanning  530021, Guangxi Zhuang Autonomous Region, China
  • Received:2010-12-15 Revised:2011-02-09 Online:2011-04-02 Published:2013-11-02
  • Contact: Jiang Hai-xing, Professor, Doctoral supervisor, Department of Gastroenterology Medicine, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China jihaxi@263.net
  • About author:Lu Zheng-feng★, Studying for master’s degree, Department of Gastroenterology Medicine, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China nnaassaa117@yahoo.com.cn
  • Supported by:

    the Natural Science Foundation of Guangxi, No. 0897008*; the “New Century Talent Project” of Guangxi, No. 2006206*

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Abstract:

BACKGROUND: Previous studies demonstrated that tumor necrosis factor-α (TNF-α) can stimulate the immunosuppression function of bone marrow mesenchymal stem cells (BMSCs), and promote hepatocyte growth factor (HGF) expression of BMSCs.
OBJECTIVE: To investigate whether BMSCs stimulated by TNF-α is an effective way to promote the expression and secretion of HGF in rat BMSCs.
METHODS: BMSCs from SD rats were isolated, cultured and purified by the whole bone marrow adherence method, generated to passage3-4 to use. TNF-α pre-stimulated BMSCs, the medium was discarded after 5 hours, and then cultured with fresh medium as experimental group, normal cultured BMSCs as blank group. The morphology of BMSCs was observed by the inverted phase contrast microscope. The expression of surface makers CD 29, CD 34, CD 44 and CD 45 of BMSCs were identified by flow cytometer. HGF mRNA and protein expression were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. HGF protein level in cell supernatants was determined by ELISA.
RESULTS AND CONCLUSION: BMSCs were isolated by the whole bone marrow adherence method with uniform and showed spindle shaped, typical swirl morphology. BMSCs expressed CD29+ (99.45%), CD34- (97.91%), CD44+ (99.52%), CD45- (98.42%); the expression level of HGF mRNA and protein and HGF in cell supernatants increased with time-dependent. The expression levels of HGF mRNA and protein and HGF in cell supernatants in experimental group were significantly higher than that in blank group (P < 0.01). BMSCs stimulated by TNF-α can effectively improve the expression and secretion of HGF.

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