Comparative Study of Culture in Vitro and Differentiate of EPCs from Bone Marrow Source of Minipig  

作　　者：Kang Cheng Guoliang Jia Haichang Wang Ronghua Luan Xiang Zhou Xiaojuan Zong 

机构地区：[1]Department of Cardiology, Xi Jing Hospital, the Fourth Military Medical University, Xi＇an,710033

出　　处：《Journal of US-China Medical Science》2006年第2期50-58,共9页美中医学（英文版）

摘　　要：Objective To compare the two methods for culturing the marrow endothelial progenitor cells （EPC） ot the minipig, elucidate the queries on separation, and provide the experiment proof for the usage of EPC transplantation in clinic. Methods We separate the mononuclear pig marrow cells following the Ficoll method. After the first screening of the cells according to their different adherent rate, we set the 24h adherent group and the 24h unadherent group. The cells are cultured in the medium containing vascular endothelial growth factor and induced differentiation. We observe the growth situation of the cells at different times and make biological evaluations, including lase confocal technique test of fluorescence labeling CD133, CD31, flk-1, and Ⅷ factor, electron microscope evaluation of ultra structure, intake test of DiI-acLDL, and measurement of secretion of the endothelial cell. Results We get a certain number of EPCs by using the two different methods, the 24h unadherent group shows a relatively homologous form, characteristic of ＂delayed proliferation＂, a high labeled rate of fluorescence, early and apparent onset of endothelial cell markers, while the markers of hematopoietic progenitor cell disappear at an early stage and express weakly, weak phagocytosis of the cell, and high amounts of NO and eNOS cell secretion. Conclusion Minipig marrow EPCs could be separated and proliferated in vitro by density gradient technique in combination with different adherent rates screening. The 24h unadherent cells are prior to 24h adherent cells and 48h unadherent cells m EPCs purity, maturity, cell function and cell numbers after induction and differentiation.

关 键 词：endothelial progenitor cells cell culture cell differentiation 

分 类 号：Q2[生物学—细胞生物学]