A novel and feasible way to cultivate and purify endothelial progenitor cells from bone marrow of children with congenital heart diseases

作     者：WU Yong-tao LI Jing-xing LIU Shuo XIN Yi WANG Zi-jian GAO Jin JI Bing-yang FAN Xiang-ming ZHOU Qi-wen 

作者机构：Department of Pediatric Cardiac Surgery Beijing Anzhen Hospital Capital Medical University Beijing 100029 China Department of Cardiac Surgery Beijing Anzhen Hospital Capital Medical University Beijing 100029 China Beijing Institute of Heart Lung and Blood Vessel Diseases Beijing Pediatric Cardiac Center （Fan XM） Beijing Anzhen Hospital Capital Medical University Beijing 100029 China Pediatric Cardiac Center Beijing Anzhen ＂FIospita＂ Capital Medical University Beijing 100029 China Institute of Biophysics of Chinese Academy of Sciences Department of Cardiopulmonary Bypass Department of Cardiopulmonary Bypass Fuwai Hospital Chinese Academy of Medical Science and Peking Union Medical College Beijing 100037 China 

出 版 物：《Chinese Medical Journal》 (中华医学杂志（英文版）)

年 卷 期：2012年第125卷第11期

页      面：1903-1907页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 08[工学] 09[农学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 071003[理学-生理学] 

基　　金：This study was supported by a grant from Science Foundation of Beijing Education Commission (No. KM200710025022) 

主　　题：congenital heart diseases endothelial progenitor cells, cell culture, bone marrow 

摘      要：Background Endothelial progenitor cells （EPCs） are used in vascular tissue engineering and clinic therapy. Some investigators get EPCs from the peripheral blood for clinic treatment, but the number of EPCs is seldom enough. We have developed the cultivation and purification of EPCs from the bone marrow of children with congenital heart disease, to provide enough seed cells for a small calibre vascular tissue engineering study. Methods The 0.5-ml of bone marrow was separated from the sternum bone, and 5-ml of peripheral blood was collected from children with congenital heart diseases who had undergone open thoracic surgery. CD34＋ and CD34＋NEGFR＋ cells in the bone marrow and peripheral blood were quantified by flow cytometry. CD34＋/VEGFR＋ cells were defined as EPCs. Mononuclear cells in the bone marrow were isolated by Ficoll density gradient centrifugation and cultured by the EndoCult Liquid Medium KitTM. Colony forming endothelial cells was detected. Immunohistochemistry staining for Dil-ac-LDL and FITC-UEA-1 confirmed the endothelial lineage of these cells. Results CD34＋ and CD34＋NEGFR＋ cells in peripheral blood were （0.07±0.05）% and （0.05±0.02）%, respectively. The number of CD34＋ and CD34＋/VEGFR＋ cells in bone marrow were significantly higher than in blood, （4.41±1.47）% and （0.98±0.65）%, respectively （P 〈0.0001）. Many colony forming units formed in the culture. These cells also expressed high levels of Dil-ac-LDL and FITC-UEA-I. Conclusion This is a novel and feasible approach that can cultivate and purify EPCs from the bone marrow of children with congenital heart disease, and provide seed cells for small calibre vascular tissue engineering.