Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (23): 4309-4316.doi: 10.3969/j.issn.1673-8225.2010.23.029

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Culture of different sources-derived endothelial progenitor cells and comparison of their biological properties

Xu Yan1, Meng Heng-xing2, Yu Zhen1, Li Chang-hong1, Qiu Lu-gui1, 2   

  1. 1Hematopathy Hospital, Institute of Hematology, Chinese Academy of Medical Sciences & Peking Union of Medical College, State Key Laboratory of Experimental Hematology, Tianjin  300020, China;
    2Tianjin Umbilical Cord Blood- Hemopoietic Stem Cell Bank, Tianjin  300384, China
  • Online:2010-06-04 Published:2010-06-04
  • Contact: Qiu Lu-gui, Master, Chief physician, Doctoral supervisor, Hematopathy Hospital, Institute of Hematology, Chinese Academy of Medical Sciences & Peking Union of Medical College, State Key Laboratory of Experimental Hematology, Tianjin 300020, China; Tianjin Umbilical Cord Blood-Hemopoietic Stem Cell Bank, Tianjin 300384, China drqiu99@medmail. com.cn
  • About author:Xu Yan, Doctor, Attending physician, Hematopathy Hospital, Institute of Hematology, Chinese Academy of Medical Sciences & Peking Union of Medical College, State Key Laboratory of Experimental Hematology, Tianjin 300020, China yanzi_zjyl@163.com
  • Supported by:

    the Natural Science Foundation of Tianjin City, No. 08JCYBJ6200*;
    the Science and Technology Innovation Special-Purpose Foundation of Tianjin City, No. 08FDZDSH03000*;
    the National Natural Science Foundation of China, No. 30871095*

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Abstract:

BACKGROUND: Compensatory response in ischemic disease included arteriogenesis and angiogenesis. Endothelial progenitor cells (EPCs) play a key role in many physiological and pathological vascular remodeling. There are still controversies in the resource and biological characteristic of EPCs.
OBJECTIVE: To isolate EPCs from peripheral blood, umbilical cord blood, bone marrow and umbilical cord, and to compare biological properties of the EPCs.
METHODS: Mononuclear cells (MNCs) of peripheral blood, umbilical cord blood, and bone marrow were isolated by density-gradient centrifugation for adherent culture. Umbilical cord-derived EPCs were made into single cell suspension by adherent culture of explants or collagenase digestion of umbilical vein for adherent culture. The obtained adherent cells were detected for cell morphology, proliferation activity, cell cycle and immunophenotype.
RESULTS AND CONCLUSION: ①Peripheral blood and umbilical cord blood-derived adherent cells grew as clones with spindle shape, and some round cells attached to them. With prolonged culture time, the round cells ablated gradually, and spindle cells had no appear proliferation. After digestion, these cells could not be subcultured. Flow cytometry showed that these cells highly expressed CD45 and partially expressed KDR. But they did not express CD31. ②Bone marrow-derived cells and umbilical cord-derived adherent cells were spindle and polygonal. Cells proliferated fast, and there was little change in cell morphology and proliferation activity after passage. Most cells were in G0/G1. Flow cytometry showed that these cells expressed neither CD14, CD45, nor CD106, HLA-DR, but highly expressed CD44, CD90, CD62E, CD73, CD95 and CD105. Some expressed endothelial marker: KDR, vWF. Umbilical cord-derived EPCs harvested by enzyme digestion were positive for CD31. Confocal microscope analysis revealed that all these cells could take up ac-LDL and bind UEA-1. Results have suggested that EPCs with different differentiation degrees and proliferation activities could be isolated from peripheral blood, umbilical cord blood, bone marrow and umbilical cord.

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