Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2011, Vol. 33 ›› Issue (3): 306-312.doi: 10.3881/j.issn.1000-503X.2011.03.019

• Original Articles • Previous Articles     Next Articles

In Vitro Effect of N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide on Differentiation from Human Adipose-derived Mesenchymal Stem Cells to Endothelial Cells

ZHAI Yu-jia1, 2, CHEN Jia1, 2, ZHANG Hong-gang2, ZHANG Jing2, ZHANG Qiu-ju2, XIU Rui-juan2   

  1. 1Graduate School, CAMS and PUMC, Beijing 100730, China
    2 Institute of Microcirculation,CAMS and PUMC, Beijing 100005, China
  • Received:2010-12-06 Revised:2011-07-01 Online:2011-07-01 Published:2011-07-01
  • Contact: ZHANG Hong-gang E-mail:zhanghg1966126@yahoo.com.cn
  • Supported by:

    the Special Social Commonweal Research Programs of the Ministry of Science and Technology(2005DIBIJ086)

Abstract: Objective To explore the effect of N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide(W7) on the differentiation from human adipose-derived mesenchymal stem cells (hADSCs) to endothelial cells. Methods hADSCs were cultured with serum-free differential medium containing 40 ng/ml vascular endothelial growth factor (VEGF) and 10ng/ml basic fibroblast growth factor (bFGF). Cells were divided into control group (differential medium without W7), high-dose group (containing 30 μmol/L W7), medium-dose group (containing 20 μmol/L W7), and low-dose group ( containing 10 μmol/L W7). The hADSCs were cultured for 8 days, and then the changes in the phenotypes of von Willebrand factor (vWF) and vessel-selective cadherin (VE-Cadherin) were detected by flow cytometry (FCM). The intracellular Ca2+ labeled with Fluo-3 was detected by laser confocal microscopy. After hADSCs planting on Matrigel, their angiogenic potentials were observed under the inverted phase contrast microscope, and the expression of extracellular regulated kinase (ERK) and phosphorylated extracellular regulated kinase (p-ERK) were evaluated by Western blot. Results After the hADSCs were cultured for 8 days, compared with the control group, the expressions of vWF and VE-Cadherin significantly increased along with the decrease of W7 level and the intracellular Ca2+ also significantly increased (P<0.01). Lumina-like vascular structure was formed in W7 treatment groups, but not in the blank control group. Compared with the blank control group, the expression of ERK showed no significant in W7 treatment groups (high-, medium-, and low-dose groups)(P>0.05); however, along with the decrease of W7 levels, the expression of p-ERK significantly increased(P<0.05). Conclusion W7 in proper levels can effectively induce the differentiation from hADSCs to endothelium by increasing the intracellular Ca2+ level and thus activating the ERK/MAPK pathway.

Key words: N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide, human adipose-derived mesenchymal stem cells, endothelial cells, induce differentiation

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