Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2012, Vol. 34 ›› Issue (3): 202-206.doi: 10.3881/j.issn.1000-503X.2012.03.002

• Original Articles • Previous Articles     Next Articles

Role of Microenvironment in the Process of Expansion of Late Endothelial Progenitor Cell in vitro

WU Li-hua1, SONG Zeng-xuan1, LIU Xu-hui2, LI Shang-zhu1, HAN Zhong-chao1, Georges Uzan2   

  1. 1Department of Internal Medicine, Institute of Hematology and Hospital of Blood Disease, CAMS and PUMC, Tianjin 300020, China
    2National Institute of Health and Medical Research,Paris 94807,France
  • Received:2012-04-05 Revised:2012-06-30 Online:2012-06-29 Published:2012-06-29
  • Supported by:

    Supported by the Technology Foundation for Selected Overseas Chinese Scholar, Ministry of Personnel of China (2006)

Abstract: Objective To study the role of the feeder layer cells as niche in the process of expansion of late endothelial progenitor cell in vitro. Methods We cultured mononuclear cells (MNC)from human peripheral blood (PB)on the plate with the feeder layer cells which were irradiated late endothelial progenitor cells(EPC)or human umbilical vein endothelial cells (HUVEC) by EGM-2. After 21 days, the numbers of obtained late EPC colonies were counted separately, and their surface antigen of the late EPC was verified by fluorescence-activated cell sorter (FACS) analysis, and their ability of forming vessel structure with Matrigel in vitro. The differentiation of single stem cell on the feeder layer cell was traced by video-microscopy. Results After 21 days of culture,(40.0±3.9)and(39.3±3.1)late EPC colonies that MNC of a handred milliliter PB were cultured, respectively, on the feeder layer cells of EPC and HUVEC were much more than (2.0±1.3) colonies cultured on without the feeder layer cells (all P <0.05). These cells also expressed CD31,CD34,eNOS,FLt-1,P1H12,Sendo,VEcadherin,and CD117, as shown by FACS analysis. Furthermore, they formed vessel structure with Matrigel in vitro. The video-microscopy showed the asymmetric cell division was participated by the feeder layer cell during the expansion of single stem cell. Conclusion The massive expansion of late EPC can be achieved by the provision of the feeder layer cells, which may be involved in the stem cell asymmetric cell division.

Key words: late endothelial progenitor cell, feeder layer cell, microenvironment, asymmetric cell division

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