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Cytotechnology

, Volume 63, Issue 3, pp 217–226 | Cite as

The characteristics of endothelial progenitor cells derived from mononuclear cells of rat bone marrow in different culture conditions

  • Nana Yang
  • Dawei Li
  • Peng Jiao
  • Bin Chen
  • Shutong Yao
  • Hui Sang
  • Mingfeng Yang
  • Jiju Han
  • Ying Zhang
  • Shucun QinEmail author
Method in Cell Science

Abstract

Endothelial progenitor cells (EPCs) derived from bone marrow are known to be heterogeneous. In this study, we tried to find favorable conditions that induce the differentiation of mononuclear cells (MNCs) from bone marrow into EPCs. The differentiation capacity of MNCs from rat bone marrow was investigated in different conditions, such as different media, different induction times and different culture surfaces. The cell morphology and endothelial biomarkers associated with differentiated MNCs were studied. Our results indicated that MNCs cultured in EGM-2MV (Endothelial cell basal medium-2, plus SingleQuots of growth supplements) developed a bursiform shape, a late EPC-like morphology, while MNCs cultured in complete medium (CM, M199 with 10% FBS, 20 ng/mL VEGF and 10 ng/mL bFGF) showed a spindle shape, an early EPC-like morphology. Cells of both morphologies were able to incorporate DiI-ac-LDL and bind lectin in vitro. MNCs cultured in EGM-2MV exhibited a higher proliferation rate and higher eNOS expression than MNCs cultured in CM. MNCs cultured in EGM-2MV had the ability to form tubes on Matrigel. Flow cytometry results indicated that CD133 expression was highest at day 12 and that the greatest number of cells positive for both FLK-1 and CD133 appeared at day 20 from cells cultured in dishes without fibronectin coating. In addition, the expression levels of CD133, CD31 and FLK-1/CD133 were not significantly different between cells of different shapes. Our experiments suggest that MNCs from bone marrow can be differentiated into late EP-like cells in EGM-2MV, which have the ability to rapidly proliferate. These MNCs can also be differentiated into early EP-like cells in CM. Additionally, fibronectin may not be necessary for the differentiation of EPCs to mature ECs after three generations. Differentiated MNCs from bone marrow in EGM-2MV have the characteristics of EPCs, although the expression levels of EPC markers were lower than previously reported.

Keywords

Endothelial progenitor cells Mononuclear cells CD133 FLK-1 

Abbreviations

MNCs

Mononuclear cells

EPCs

Endothelial progenitor cells

eNOS

Endothelial nitric oxide synthase

FCM

Flow cytometry

ac-LDL

acetylated low density lipoprotein

Notes

Acknowledgments

This study was supported by the National Natural Science Foundation of China (Grant No. 30971098) and the Province Natural Science Foundation of Shandong (Grant No. Z2008C03).

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Nana Yang
    • 1
  • Dawei Li
    • 1
  • Peng Jiao
    • 1
  • Bin Chen
    • 1
  • Shutong Yao
    • 1
  • Hui Sang
    • 1
  • Mingfeng Yang
    • 1
  • Jiju Han
    • 1
  • Ying Zhang
    • 1
  • Shucun Qin
    • 1
    Email author
  1. 1.Institute of AtherosclerosisTaishan Medical UniversityTaianChina

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