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Culture and proliferation of human umbilical cord blood endothelial progenitor cells in vitro

SHAO Chun-yi, CHEN Jun-zhao, FU Yao, CHEN Ping, LU Wen-juan, FAN Xian-qun   

  1. Department of Ophthalmology, the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
  • Online:2013-10-28 Published:2013-10-31
  • Supported by:

    National Natural Science Foundation of China, 31000443; Shanghai Science and Technology Committee Foundation, 11JC1407000; Shanghai Pujiang Talent Program, 09PJ1407200; Shanghai Jiaotong University School of Medicine Doctor Innovation Foundation, BXJ201227

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Abstract:

Objective To investigate the feasibility of human umbilical cord blood endothelial progenitor cells (EPCs) in substitute for corneal endothelial cells as seed cells. Methods Mononuclear cells were isolated from human umbilical cord blood by density gradient centrifugation, and were plated in endothelial growth medium on fibronectin coated culture dish with EGM-2 as culture fluid. The ability of EPCs in uptaking acetylated low density lipoprotein (AcLDL) and binding Ulex Europaeus Lectin 1 (UEA1) was determined, and the expression of CD133, CD34 and vWF was detected by immunofluorescence method. Results The human umbilical cord blood EPCs exhibited polygonal or short spindle shape. Fibronectin-coated dish facilitated the adhesion of EPCs. Cellular morphology and proliferation ability were not changed by passage. The human umbilical cord blood EPCs could uptake AcLDL, bind UEA1, and express CD133, CD34 and vWF. Conclusion Human umbilical cord blood EPCs may be favorable candidate in replacement of corneal endothelial cells as seed cells.

Key words: human umbilical cord blood, endothelial progenitor cells, cornea, endothelial cells