菲立磁标记大鼠骨髓基质干细胞对其生物活性的影响

Biological characteristics of Feridex labeled bone marrow stem cells

目的观察菲立磁（Feridex）标记对大鼠骨髓基质干细胞（BMSCs）的增殖及向肝细胞分化能力的影响，探讨Feridex标记大鼠BMSCs的最佳标记方案。方法sD大鼠BMSCs按Feridex浓度11．2、14．0、16．8、19．6mg／L分组并进行标记，设空白对照组（无Feridex标记的BMSCs）。采用普鲁士兰染色和透射电镜鉴定Feridex标记后各组BMSCs的标记效率。噻唑蓝（MTT）比色法检测Feridex标记后各组BMSCs的增殖水平。逆转录-聚合酶链反应（RTPCR）检测Feridex标记后各组BMSCs经肝细胞生长因子（HGF）、表皮细胞生长因子（EGF）共同诱导后ALB和AFP基因的表达水平。结果11．2、14．0、16．8、19．6mg／L组的标记率分别为71％、83％、91％、96％。16．8mg／L和19．6mg／L组的标记率显著高于11．2mg／L和14mg／L组（P〈0．05）。11．2、14．0、16．8mg／L组的细胞增殖水平及诱导后ALB、AFP基因表达水平与空白对照组比较差异无统计学意义（P〉0．05），而19．6mg／L组的细胞增殖水平及诱导后ALB、AFP基因表达水平显著低于空白对照组（P〈0．05）。结论Feridex浓度16．8mg／L可能是Feridex标记BMSCs最适标记浓度，该浓度既能使Feridex对BMSCs的标记达到较理想效果，同时不影响BMSCs的增殖及向肝细胞分化的能力。

Objective To study the effects of Feridex labeling on bone marrow stem cells （BMSCs） ability to proliferate and differentiate into hver cells, searching for the optimal schedule of Feridex la- beling. Methods BMSCs were labeled by Feridex at different concentration gradient, including 11.2, 14.0,16.8,19.6 mg/L. Blank control group （ no Feridex labeling） was set up. The labeling efficiency was detected by Prussian blue staining and electron microscope ,and proliferating ability was evaluated by MTI＂ methods. Labeled cells were transdifferentitated into hepatic cells by hepatocyte growth factor （HGF） and epidermis growth factor （EGF）. The expression levels of ALB and AFP mRNA were analyzed semiquantitatively by using reverse transcription polymerase chain reaction （RT-PCR） techniques. Results The la- beling efficiency of 11.2,14.0,16.8,19.6 mg/L groups was 71%, 83 % ,91% ,96 %, respectively. The la beling efficiency of 16.8 mg/L and 19.6 mg/L groups was significantly greater than that of 11.2 mg/L and 14 mg/ml groups （ P 〈 0.05）. There was no significant difference in the abilities of prohferation of BMSCs and the expression levels of ALB and AFP mRNA of transdifferentitated cells among 11.2,14.0, 16.8 mg/L groups and blank control group （ P 〉 O. 05 ） , while the indexes of 19.6 mg/ml group were low- er than those of blank control group statistically （ P 〈 0.05 ）. Conclusion The labehng efficiency was high and the ability of BMSCs＇ proliferation and differentiation into liver cells was not affected at 16.8 mg/L of Feridex,so it may be the most suitable concentration for labeling.