›› 2011, Vol. 31 ›› Issue (7): 913-.doi: 10.3969/j.issn.1674-8115.2011.07.010

• Original article (Basic research) • Previous Articles     Next Articles

Effects of hypoxia pathway on multipotential differentiation of bone marrow stromal stem cells

ZENG Wen1, ZHANG Wei1, WANG Jun1, DENG Ruo-xian2, ZHOU Qi1, WEI Li1, QI Jin1, DENG Lian-fu1   

  1. 1.Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Institute of Traumatology and Orthopedics, Shanghai Key Laboratory of Combination of Traditional Chinese and Western Medicine in Prevention and Therapy of Osteo-arthropathy, Shanghai 200025, China；2.School of Medical Devices and Food Sciences, University of Shanghai for Science and Technology, Shanghai 200093, China
  • Online:2011-07-28 Published:2011-07-27
  • Supported by:

    National Natural Science Foundation of China, 30872641；Shanghai Science and Technology Committee Foundation, 09DZ2200500,10DZ2211500；Technology Fund of Shanghai Jiaotong University School of Medicine, YZ1029

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Abstract:

Objective To investigate the effects of hypoxia-inducible factor 1α(HIF-1α) on the multipotential differentiation of bone marrow stromal stem cells (MSCs) in the hypoxia pathway. Methods Conditional gene knockout of VHL gene of MSCs from transgenic mice was performed with Ad-Cre (gene knockout group), and control group was established (MSCs from transgenic mice infected with Ad-GPF). The expression of HIF-1α mRNA was detected by Real-Time PCR. Chondrogenic culture and osteogenic culture of MSCs were conducted for 14 d in two groups, and osteogenic culture of MSCs was conducted for 21 d in two groups, with culture of MSCs under 5% O2 in gene knockout group and culture of MSCs under 20% O2 in control group. The expression of chondrocyte marker of typeⅡcollagen (ColⅡ), adipocyte marker of peroxisome proliferator-activated receptors gamma (PPARγ) and osteoblast markers of osteocalcin (OC) and alkaline phosphatase (ALP) mRNA was detected by Real-Time PCR. The distributions of positive cells with ColⅡ staining and ALP staining were observed by light microscopy in two groups. Results The expression of HIF-1α mRNA in gene knockout group was significantly higher than that in control group (P<0.05). The expression of ColⅡ, PPARγ, OC and ALP mRNA in gene knockout group cultured under 5% O2 was significantly higher than that in control group cultured under 20% O2 (P<0.05). Compared with control group, the numbers of positive cells with ColⅡ staining and ALP staining were larger. Conclusion HIF-1α can promote the differentiation of MSCs into chondrocytes, adipocytes and osteoblasts under 5% O2.

Key words: hypoxia-inducible factor 1α, bone marrow stromal stem cells, gene knockout, differentiation