摘要: |
目的 采用拟水平均匀实验设计,探讨超声联合微泡促人骨髓间充质干细胞分泌SDF-1的最优参数组合。方法 选定超声辐照强度、超声辐照时间、微泡浓度三个因素,又分别按超声辐照强度 0 (对照)、0.2、0.4、0.6、0.8 W/cm2,辐照时间 0 (对照)、20、30、40、60 s,微泡浓度 0 (对照)、102、104、106、108 N/ml,每因素五个水平设计均匀设计表,根据均匀设计表U15(155) 进行超声辐照,辐照后细胞继续培养24 h,然后检测细胞存活率及细胞培养上清液中的SDF-1浓度。以较高的SDF-1浓度和较低的细胞死亡率为目标,应用多元线性回归和逐步回归分析确定最佳参数配比,并对统计分析后的优化参数再进行实验验证。结果 综合考虑各因素间关系及回归分析结果后,确定超声联合微泡促人骨髓间充质干细胞分泌SDF-1的最优参数组合为辐照强度 0.6 W/cm2,辐照时间 30 s,微泡浓度 106/ml,验证实验显示在此条件下超声辐照后 SDF-1分泌量达 551.67±40.88 pg/ml,细胞存活率达 88.51±4.03%。结论 应用拟水平均匀设计可有效优化参数条件,使超声辐照后间充质干细胞的SDF-1分泌量和细胞存活率达到相对匹配的数值。 |
关键词: 均匀设计 超声辐照 微泡 骨髓间充质干细胞 SDF-1 |
DOI: |
投稿时间:2021-04-29修订日期:2021-05-24 |
基金项目:重庆市自然科学基金(cstc2018jcyjAX0011) |
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Optimization of ultrasound combined with microbubbles promote SDF-1 expression in human mesenchymal stem cells according to the quasi-level uniform design |
Li Lu,He Fen,Yang Feng Wu,Gu Jing,Xu Ya Li |
(General Hospital of Western Theater Command) |
Abstract: |
Objective To investigate the optimal parameters of the ultrasound combined with microbubbles promoting SDF-1 expression in human mesenchymal stem cells according to the quasi-level uniform design. Methods The acoustic intensity, irradiation time, and the dosage of MBs were selected, then five levels of each factor were chosen, acoustic intensity (0 W/cm2、0.2 W/cm2、0.4 W/cm2、0.6 W/cm2、0.8 W/cm2),irradiation time (0 s、20 s、30 s、40 s、60 s), and the dosage of MBs (0 N/ml、102 N/ml、104 N/ml、106 N/ml、108 N/ml). The uniform design table U15(155) and the using tables were set, and the rat mesenchymal stem cells were radiated according to the uniform experimental design. After ultrasound radiation, the cells were cultured for 24 h, then the concentration of SDF-1 in cultural supernatant and the cell viability were detected. Following the principle of more SDF-1 secretion and less cell death, the optimized parameters were determined by regression analysis and then validated by orthogonal experimental. Results Considering good cell viability, the optimal conditions for SDF-1 secretion were acoustic intensity of 0.6 W/cm2 , irradiation time of 30 s and the dosage of MBs of 106 N/ml. The results of verification experiments showed that after microbubble mediated ultrasound irradiation, the concentration of SDF-1 was 551.67±40.88 pg/ml and the cell viability was 88.51±4.03% by using the optimized parameters. Conclusion The application of quasi-level uniform design can effectively optimize the parameters of microbubble mediated ultrasound irradiation, and find a balance between SDF-1 secretion and cell viability of mesenchymal stem cells. |
Key words: Uniform design Ultrasonic radiation Microbubble Mesenchymal stem cell SDF-1 |