Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (6): 1000-1004.doi: 10.3969/j.issn.1673-8225.2011.06.012

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Co-culture, proliferation and migration of human umbilical cord blood-derived endothelial progenitor cells with vascular endothelial growth factor and basic fibroblast growth factor 

Zhou Jian-liang, Zou Ming-hui, Chen Yi-chu, Zhou Cheng, Shi Jia-wei, Dong Nian-guo   

  1. Department of Cardiovascular Surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan  430022, Hubei Province, China
  • Received:2010-09-25 Revised:2010-11-19 Online:2011-02-05 Published:2011-02-05
  • Contact: Dong Nian-guo, Professor, Doctoral supervisor, Department of Cardiovascular Surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China zjldjl@sina.com
  • About author:Zhou Jian-liang☆, Studying for doctorate, Attending physician, Department of Cardiovascular Surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China zhoujianliang2010@163.com
  • Supported by:

    the National High Technology Research and Development Program of China (“863” Program), No. 2009AA03Z420*; National Natural Science Foundation of China, No. 30872540*; Chenguang Plan of Wuhan, No. 200950431177*

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Abstract:

BACKGROUND: At present seeded cells of re-endothelialization for tissue engineered heart valve (TEHV) are mainly from mature endothelial cells; endothelial progenitor cells (EPCs) are precursor cells of endothelial cells, which attracts more and more people’s attention.
OBJECTIVE: To isolate and proliferate human umbilical cord blood-derived (HUCB) endothelial progenitor cells (EPCs), and to observe their biological characteristics in vitro.
METHODS: Mononuclear cells (MNCs) were isolated from fresh umbilical blood by density gradient centrifugation. MNCs were proliferated and cultured in medium supplemented with vascular endothelial grow factor (VEGF) and basic fibroblast growth factor (bFGF). Attached cells were identified by morphology, immunofluorescence staining and flow cytometry. And the EPCs were compared with human umbilical vein endothelial cells (HUVECs) in the capacity of the proliferation and migration.
RESULTS AND CONCLUSION: With the prolongation of culture and induction time, the morphology of attached cells were significantly changed from small round cells to spindle cells, and gradually differentiated into typical mature endothelial cell cobblestone-like morphology, which could form characteristic clone. After 7 days induction in vitro, more than 90% of attached cells presented both Dil-ac-LDL and FITC-UEA-I positive. On day 7, flow cytometric analysis showed that the positive staining rate of attached cells for VEGFR2, CD34 and CD133 were (77.4±4.9)%, (52.4±6.6)%, and (19.4±2.1)%, respectively. On day 28, flow cytometric analysis showed that the positive staining rate of attached cells for VEGFR-2 and CD34 were (81.1±7.4)% and (7.6±3.1)%, but CD133 expression was not detected. The capacity of proliferation and migration of EPCs were significantly higher than that in HUVECs (P < 0.05), and the total number of cells reached 109 /L. The results showed that EPCs can be isolated and purified from HUCB by density gradient centrifugation and adherence screening method. EPCs can be induced and differentiated into endothelial cells, with high capacity of proliferation and migration.

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