摘要
目的:研究小鼠骨髓来源内皮祖细胞(EPCs)的体外培养方法及体外分化特征。方法:采用梯度离心法分离小鼠骨髓单个核细胞并进行诱导培养,用流式细胞术和免疫细胞化学对细胞表型特征进行检测,分别采用MTT和Transwell方法对其增殖、迁移能力进行检测。结果:体外培养4 d至14 d,CD133、SCA-1及CD117的表达率分别从11.05%、29.32%及16.45%降至2.29%、7.82%及3.92%;而VEGFR-2、CD31、VE-cadherin、eNOS及vWF的表达率分别从12.21%、8.43%、18.27%、7.11%及32.61%增加至62.13%、32.96%、67.73%、27.89%及82.16%;吞噬acLDL和结合UEA-I的细胞从57.18%增加至86.70%。在0μg/L至80μg/L VEGF浓度梯度下,细胞增殖率和迁移率分别增加58.03%和33.83%。结论:EGM-2内皮培养基可稳定培养小鼠骨髓来源EPCs,培养过程中其干细胞标志物表达逐渐降低,而内皮细胞标志物表达及特性不断增加,VEGF可显著促进其增殖和迁移。
AIM：To investigate the culture method and differentiation characteristics of endothelial progenitor cells（EPCs） derived from mouse bone marrow.METHODS： Bone marrow mononuclear cells were isolated from BALB/c mice by density gradient centrifugation and cultured in fibronectin-coated dishes with EGM-2 medium.The phenotypic characteristics were determined by immunofluorescence and fluorescence-activated cell sorter.The proliferation and migration capacities were detected by MTT and Transwell assays,respectively.RESULTS： While the cells were cultured in vitro,the expression of CD133,SCA-1 and CD117 between day 4 and day 14 decreased from 11.05%,29.32% and 16.45% to 2.29%,7.82% and 3.92%,respectively.Nevertheless,VEGFR-2,CD31,VE-cadherin,eNOS and vWF increased from 12.21%,8.43%,18.27%,7.11% and 32.61% to 62.13%,32.96%,67.73%,27.89% and 82.16%,respectively.The capacities of uptaking acLDL and binding UEA-I increased from 57.18% to 86.70%.Under the concentration gradient of VEGF from 0 μg/L to 80 μg/L,the capacities of proliferation and migration increased by 58.03% and 33.83% respectively.CONCLUSION： EPCs derived from mouse bone marrow can be cultured in EGM-2 medium supplemented with proper growth factors.With the differentiation,EPCs loss their stem cell markers and express endothelial specific markers gradually.VEGF plays an important role in promoting proliferation and migration of EPCs in a dose-dependent manner.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2011年第6期1121-1126,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81071534)
关键词
内皮祖细胞
体外培养
分化特征
Endothelial progenitor cells
Culture in vitro
Differentiation characteristics
