Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2012, Vol. 34 ›› Issue (6): 539-544.doi: 10.3881/j.issn.1000-503X.2012.06.001

• Original Articles •     Next Articles

Differentiation of Human Embryonic Stem Cells to Endothelial Cells via Improved Three-dimension Approach

SU Wei-jun1, WANG Bao-yu2, SONG Xiang-he3, WANG Li-na4, LIU Yan-hua4,ZHOU Man-qian4, TONG Ling-ling5, LI Zong-jin5   

  1. 1Department of Pathology, Nankai University School of Medicine, Tianjin 300071, China;2Department of Endocrinology, Nankai University People's Hospital, Tianjin 300121, China;3Department of Cardiology, Rizhao Hospital of Traditional Medicine, Rizhao, Shandong 276800,China;4Department of Immunology, Nankai University School of Medicine, Tianjin 300071, China;5Department of Pathophysiology, Nankai University School of Medicine, Tianjin 300071, China
  • Received:2012-01-13 Online:2012-12-30 Published:2012-12-30
  • About author:WANG Bao-yu Tel: 022-27557477, E-mail:wbylwg@sina.com;LI Zong-jin Tel:022-23509475, E-mail: zongjinli@nankai.edu.cn
  • Supported by:
    Supported by the National Natural Sciences Foundation of China(31071308) and Tianjin Research Program of Application Foundation and Advanced Technology (12JCZDJC24900)

Abstract: Objective To establish an improved three-dimension (3D) and serum-free approach to differentiate human embryonic stem cells (hESCs) into endothelial cells, and detect the endothelial functions of the obtained cells. Methods We cultured undifferentiated H9 human embryonic stem cell line in low-adhesion dishes to form embryonic bodies (EBs). After 12 days, EBs were harvested, re-suspended into rat tail collagen type I, and put into the incubator (37℃). After 30 minutes, EGM-2 culture medium was added to the solidified collagen, and the EBs were cultured for another 3 days to form embryonic body-sproutings (EB-sproutings). EB-sproutings were digested with 0.25% collagenase I and 0.56 U/ml Liberase Blendzyme for 20 minutes respectively, and the CD31+ cells were sorted by FACS. The endothelial functions were tested by Dil-ac-LDL uptake assay and tube formation assay. Results This approach raised the efficiency of endothelial differentiation to 18%, and also avoided the contamination with animal materials. The obtained hESC-derived endothelial cells (hESC-ECs) had the similar pattern of surface biomarkers as human umbilical vein endothelial cells (HUVECs), and their endothelial functions were confirmed by the uptake of Dil-ac-LDL and the tube formation on Matrigel. Conclusions The improved 3D approach can enhance the efficiency of differentiation from hESCs into endothelial cells. Furthermore, serum free differentiation system may be applied in future hESC-based therapies for various ischemic diseases.

Key words: human embryonic stem cells, human embryonic stem cell-derived endothelial cells, three dimension differentiation, CD31

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