小鼠骨髓内皮祖细胞培养及磁标记研究

Culture and Magnetically Labeling of Endothelial Progenitor Cells from Mouse Bone Marrow

目的建立一种小鼠骨髓单个核细胞体外诱导培养内皮祖细胞(EPCs)的方法,并探讨用多聚赖氨酸包被的氧化铁纳米粒子(Fe2O3-PLL)对其进行标记的可行性。方法利用差时贴壁法分离骨髓单个核细胞,诱导培养得到EPCs,利用免疫组织化学、荧光染色对细胞特性进行鉴定;利用Fe2O3-PLL标记细胞,并用普鲁氏蓝染色法对其鉴定,MTT法评价Fe2O3-PLL对细胞生长的影响。结果差时贴壁法分离小鼠骨髓单个核细胞诱导培养呈内皮细胞形态,表达CD31、CD34、vWF,Dil-ac-LDL、FITC-UEA-1双荧光染色阳性;Fe2O3-PLL对细胞增殖无明显影响。结论该方法可高效培养扩增内皮祖细胞;Fe2O3-PLL可高效标记该细胞。

Objective To establish a method for isolation and culture of mouse bone marrow-derived endothelial progenitor cells（EPCs） and to detect the feasibility of magnetically labeling this cells. Methods Mononuclear cells were isolated from mouse bone marrow, cultured inductively for EPCs. The EPCs were identified by immunochemistry and fluorescence staining. The EPCs were labeled with Fe2O3-PLL and the intracellular iron was identified with prussian blue staining. MTT assay was used to evaluate proliferation of Fe2O3-PLL labeled EPCs. Results These cells cultured in vitro expressed EPCs-specific antigens, such as CD31, CD34 and vWF, and were shown to endocytose Dil- ac-LDL and FITC-UEA-1. Magnetically labeled by Fe2O3-PLL, the proliferation of EPCs was not affected significantly. Conclusion An efficient method for culture of EPCs was established; the EPCs can be labeled with Fe2O3-PLL efficiently.