利用SPR生物传感器实时监测干细胞促血管新生旁分泌因素的变化  

Real - time monitoring of paracrine secreting factors of stem cells by SPR biosensors

作　　者：王晖 高斌[2] 潘杰[2] 程志俭[2] WANG Hui;GAO Bin;PAN Jie;CHENG Zhijian(Department of General Surgery,Shanghai Xuhui Center Hospital,Shanghai 200031,China;Department of Vascular Surgery,the Fifth People＇s Hospital of Shanghai Affiliated to Fudan University,Shanghai 200240)

机构地区：[1]上海市徐汇区中心医院普外科,上海200031 [2]复旦大学附属上海市第五人民医院血管外科,上海200240

出　　处：《徐州医科大学学报》2018年第8期535-539,共5页Journal of Xuzhou Medical University

基　　金：上海市闵行区卫计委资助项目（2013MW02）

摘　　要：目的探讨用表面等离子体共振（surface plasmon resonance，SPR）技术生物传感器实时定量和定性监测干细胞分化中旁分泌血管内皮生长因子（vascular endothelial growth factor，VEGF）的时相性变化及灵敏度。方法通过样品与SPR传感器芯片表面修饰的VEGF单抗特异性识别引起金膜表面折射率变化，进而导致SPR角的变化，实现实时监测体外促骨髓间充质干细胞定向分化内皮细胞体系中VEGF的变化。结果SPR传感器可以监测MSC旁分泌的微量VEGF，在分化培养第5天明显增高，第6天达到峰值，检测下限可达2nmol／L。结合速率常数为4．68×10-5／ms，解离速率常数为1．73×10-4／s，解离速率平衡常数为3．71×10-10mol／L。结论SPR传感器能定性、定量检测干细胞在定向分化过程的旁分泌细胞因子，与传统方法相比，SPR技术具有较高的灵敏度、准确性且操作简便。Objective To explore the real - time quantitative and qualitative monitoring of the time - dependent changes and sensitivity of vascular endothelial growth factor （VEGF） in the process of stem cell differentiation by surface plasmon resonance （SPR） biosensors. Methods The detection of VEGF in the process of MSC differentiation was based on the changes of SPR angles, which was subsequently caused by the changes in the refractive index of gold film induced by the specific recognition of samples by VEGF antibody modified on the surface of SPR chips. Results The level of VEGF was detected by SPR biosensors, which significantly increased from Day 5 and peaked on Day 6. The limit of detection （LOD） was 2 nmo]/L. The binding rate constant was 4.68 × 105/ms, the dissociation rate constant was 1.73 × 10 -4/s, and the dissociation rate equilibrium constant was 3.71 × 10 -10 mol/L. Conclusions SPR biosensors can qualitatively and quantitatively detect the paracrine cytokines of stem cells in the process of differentiation. Compared with the traditional methods, SPR is advantageous in high sensitivity, high accuracy and easy operation.

关 键 词：生物传感器 干细胞 旁分泌 血管内皮生长因子 

分 类 号：R654.4[医药卫生—外科学]