不同浓度菲立磁对大鼠间充质干细胞标记效率和细胞活力的影响

目的探讨不同浓度菲立磁标记大鼠骨髓间充质干细胞（MSCs）的磁标记效率及对细胞生长活力的影响，寻找最佳标记浓度。材料与方法菲立磁与多聚左旋赖氨酸（PLL）混合制备菲立磁-PLL复合物。将不同浓度的菲立磁-PLL复合物与培养基混合（铁的终浓度分别为150μg／ml、100μg／ml、50μg／ml、25μg／ml、10μg／ml和5μg／ml），加入MSCs孵育过夜。分别于标记后1天、1周、2周、3周、4周行铁染色、铁含量测定及细胞活力检测。结果菲立磁-PLL标记组细胞铁含量显著高于单纯菲立磁标记组（P＜0．05），25μg／ml浓度组细胞铁含量显著高于10μg／ml及其以下浓度组（P＜0．05），但与50μg／ml及其以上浓度组差异无统计学意义（P＞0．05）。台盼蓝排除试验显示，100μg／ml及其以下浓度组细胞活力与对照组之间差异无统计学意义（P＞0．05）。结论以25μg/ml铁浓度标记干细胞不仅标记效率高，而且对细胞活力无明显影响．为菲立磁标记MSCs的最佳浓度。

A Study on Labeling Efficiency and Cellular Viability for Rat Mesenchymal Stem Cells Labeled with Different Concentration of Feridex

Objective To explore the labeling efficiency and cellular viability of rat mesenchymal stem cells labeled with different concentration of feridex, and find an optimal concentration of feridex for cells magnetic labeling. Materials and Methods Feridex was mixed with poly left lysine (PLL) to obtain a complex of feridex PLL. The culture media containing feridex PLL of different concentration were added to the MSCs, the ultimate concentration of feridex was 150 ug/ml, 100 ug/ml, 50 ug/ml, 25 ug/ml, 10 ug/ml, and 5 ug/ml respectively. After incubation overnight, prussian blue staining, atomic absorption spectrophotometry, and trypan blue exclusion test were performed at 24 h, 1 W, 2 W,3 W, 4 W respectively after labeling.Results The iron content for feridex PLL labeling group was higher than for the control group (P<0.05). In the feridex PLL groups, the iron content of the 25ug/ml group was higher than of the 10ug/ml or below groups (P<0.05), but not lower than of the 50ug/ml or above groups. Trypan blue exclusion test showed the viability of the cells labeled with feridex of 100 ug/ml or below concentration was not significantly impaired at different time. Conclusion The iron concentration of 25 ug/ml is efficient and safe, can be considered an optimal dose for cells labeling with feridex.