超顺磁性氧化铁标记人Flk-1^+CD31^-CD34^-间充质干细胞对细胞增殖及向神经细胞分化的影响  

Effects of superparamagnetic iron oxide on proliferation and neural differentiation of human mesenchymal stem cells

作　　者：王常珺[1] 卢姗[2] 冯铭[1] 韩钦[2] 魏俊吉[1] 王任直[1] 赵春华[2] 

机构地区：[1]北京协和医院神经外科 [2]中国医学科学院基础医学研究所组织工程中心,北京100005

出　　处：《基础医学与临床》2008年第9期930-935,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30571901);国家高技术研究发展计划(国家863计划:2006AA02A115)

摘　　要：目的探讨磁共振增强剂超顺磁性氧化铁(SPIO)标记人Flk-1+CD31-CD34-间充质干细胞(hMSCs)对细胞增殖及向神经细胞分化的影响。方法实验组hMSCs与含SPIO50mg/L及多聚赖氨酸(PLL)1.5mg/L的培养基孵育过夜(12~18h)。通过生长曲线测定、台盼蓝染色及流式细胞仪检测评价SPIO对hMSCs增殖能力、细胞活性及细胞表面标志物的影响。经神经诱导后,通过免疫荧光法测定神经细胞特异性标志物的表达。结果连续测定7d细胞活性,活细胞率均超过90%,对照组及实验组生长曲线符合对数生长模式。两组细胞均表达CD44、CD105及Flk-1,而CD31、CD34为阴性。神经诱导后,神经特异性标志物的荧光A值在两组间无统计学差异。结论SPIO作为磁共振活体示踪剂是安全、有效的。Objective To label Flk-1 ^＋ CD31^- CD34^- human mesenchymal stem cells （hMSCs） with superparamagnetic iron oxide （SPIO） and to evaluate the effect of SPIO on proliferation and neural differentiation of labeled cells. Methods hMSCs were incubated with SPIO （50 mg/L） and PLL （ 1.5 mg/L） overnight（ 12 - 18 hours）. Both labeled and unlabeled cells went through growth curve test, Trypan blue staining and flow cytometer to evaluate the effects of SPIO on cell proliferation, cell viability and surface markers. Immunofluorescence assay was conducted for neuron and neuroglia specific cell surface markers after neural induction protocols were used. Results Cell viability of the two groups were both more than 90% for 7 days. There was no significant difference in cell viability and growth curve test between two groups. The results of flow cytometer showed that both labeled and unlabeled cells expressed CD44, CD105 and Flk-1 markers, while CD31 and CD34 were negative. After neural induction, the statistical analysis of A value for all the markers showed no significant difference between the two groups. Conclusion SPIO, as MRI cellular contrast, is safe and efficient.

关 键 词：超顺磁性氧化铁 间充质干细胞 干细胞移植 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R445.2[医药卫生—基础医学]