Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (40): 7450-7454.doi: 10.3969/j.issn.1673-8225.2011.40.010

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Culture, large-scale expansion and biocompatibility of endothelial colony forming cells

Chen Chao, Yang Shu-hua, Feng Yong, Chen Dong, Yu Qian, Wang Xiao-hong   

  1. Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan  430022, Hubei Province, China
  • Received:2011-03-02 Revised:2011-04-06 Online:2011-10-01 Published:2011-10-01
  • Contact: Yang Shu-hua, Professor, Doctoral supervisor, Chief physician, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China shuhuayabc@vip.sina.com
  • About author:Chen Chao☆, Studying for doctorate, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China chenchao027@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30973044*

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Abstract:

BACKGROUND: There is still uncertain about in vitro culture and expansion of endothelial colony forming cells (ECFCs) and its application in tissue engineering.
OBJECTIVE: To elucidate a method for in vitro culture, identification and large-scale expansion of ECFCs and to investigate its biocompatibility with biological material.
METHODS: Monocytes were isolated by Ficoll density gradient centrifugation from human umbilical cord blood, in vitro cultured and expanded in EGM-2 medium on tissue culture plates precoated with type I rat tail collagen. Then cells were characterized by immunofluorescence staining. The growth kinetics, tube formation capacity and biocompatibility with nano-hydroxyapatite/tricalcium phosphate (HA/TCP) were observed.
RESULTS AND CONCLUSION: During culture, the cells displayed cobble-stone morphology with outgrowth, took up DiI-Ac-LDL, bound FITC-UEA-1, and stained positively for cell markers CD31, vWF, KDR and Tie-2. These cells demonstrated high proliferation rate after passage, formed closed network structures on Matrigel and the bioactivity remained stable when co-cultured with HA/TCP. Results indicate it is feasible to isolate and large-scale expand ECFCs from human umbilical cord blood in vitro and ECFCs may serve as seed cells in tissue engineering.

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