Chinese Journal of Tissue Engineering Research ›› 2017, Vol. 21 ›› Issue (9): 1444-1449.doi: 10.3969/j.issn.2095-4344.2017.09.024

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Runx2 over-expression effect on expression of osteogenesis related genes in osteogenic differentiation of human umbilical cord blood mesenchymal stem cells

Zhou Da-kai1, Li Hui-ning1, Ma Shan-shan2, Cheng Tian3   

  1. 1Department of Spinal Surgery, Xinxiang Central Hospital, Xinxiang 453000, Henan Province, China; 2School of Life Science, Zhengzhou University, Zhengzhou 450001, Henan Province, China; 3First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
  • Online:2017-03-28 Published:2017-03-31
  • Contact: Cheng Tian, M.D., Attending physician, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
  • About author:Zhou Da-kai, Master, Attending physician, Department of Spinal Surgery, Xinxiang Central Hospital, Xinxiang 453000, Henan Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. U1404313

Abstract:

BACKGROUND: Runx2 plays a central role in osteogenic differentiation, which is of important significance in new bone formation.
OBJECTIVE: To observe the effect of Runx2 over-expression on osteogenic differentiation of human umbilical cord blood mesenchymal stem cells.
METHODS: Runx2 was generated by RT-PCR and the recombinant adenovirus (pAd-Runx2) was constructed. The viral titer was determined by air dilution method. After being transfected into human umbilical cord blood mesenchymal stem cells, the green fluorescence was observed under fluorescence microscope. Real-time fluorescent quantitative PCR and western blot were used to detect mRNA expression changes of osteogenesis related genes, Runx2, OCN, BMP-2, ALP, in human umbilical cord blood mesenchymal stem cells at 1, 3, 7 and 14 days after transfection.
RESULTS AND CONCLUSION: The recombinant adenovirus was successfully constructed and the titer was 1.7×1010 pfu/L. After infection by pAd-Runx2, human umbilical cord blood mesenchymal stem cells expressed green fluorescence protein clearly under the fluorescence microscope. Cells in the transfected group differentiated into osteoblast-like cells, and those in the control group stayed the same as pre-infected. The expression of Runx2, OCN, BMP-2 and ALP in the transfected group increased over time to some extent, but these changes were not detected in the control group. These findings indicate that Runx2 over-expression can promote the osteogenic differentiation of human umbilical cord blood mesenchymal stem cells.

 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

Key words: Cord Blood Stem Cell Transplantation, Core Binding Factor Alpha 1 Subunit, Adenoviridae, Tissue Engineering

CLC Number: