聚乙烯亚胺包被的Fe_3O_4磁纳米颗粒对hADSCs生物学特性的影响

Effects of polyethylenimine covered Fe_3O_4 superparamagnetic iron oxide on human adipose mesenchymal stem cells

目的探讨聚乙烯亚胺(PEI)包被的超顺磁性氧化铁(SPIO)对人脂肪间充质干细胞(hADSCs)生物学特性的影响。方法从人皮下脂肪组织中分离、培养hADSCs并鉴定,选取第3代hADSCs,分别用含铁浓度为6、8、10和12 mg/L的DMEM/F12培养液共孵育。通过普鲁士蓝染色、MTT细胞增殖实验、锥虫蓝染色及多向分化诱导实验来探讨PEI-SPIO标记后,hADSCs的标记能力、细胞增殖和分化能力。结果 1)当铁终浓度为8、10和12 mg/L时,PEI-SPIO对hADSCs的标记率为99%,当铁终浓度<8 mg/L时,PEI-SPIO对hADSCs的标记率为96%。2)当铁终浓度为6和8 mg/L时,PEI-SPIO标记的hADSCs的活细胞比率为99%,而当铁终浓度为10和12 mg/L时,活细胞比率为(79.03±2.25)%和(70.23±1.36)%,显著低于对照组(P<0.01)。3)当铁浓度为10和12 mg/L时,在诱导hADSCs分化过程中,细胞绝大多数死亡。结论铁浓度8 mg/L为PEI-SPIO标记的最适宜浓度,既能高效的标记hADSCs,又不影响hADSCs的细胞增殖、多向分化能力等生物学特性。

Objective To investigate the effects of superparamagnetic iron oxide covered by polyethylenimine （PEI） on human adipose mesenchymal stem cells （hADSCs）. Methods The adipose tissue comes from human subcutane- ous tissue. The third passage of hADSCs were divided into four groups, and incubated in DMEM/F12 containing PEI-SPIO of 6,8,10,12 mg/L. Detected by Prussian blue staining, Trypan blue staining, MTT test and inductions of muhi-directional differentiation to evaluate the ability of SPIO on cell labeling efficiency, cell viability, proliferation and differentiation. Results 1 ） The labeling efficiency of SPIt （8,10,12 mg/L） was 99%. However the labeling efficiency of SPIO （6 mg/L）was 96%. 2）In 6,8 p.g/mL group, the cell survival rate was 99%. But in 10 mg/L and 12 mg/L group, the cell survival rate was （79.03 ± 2. 25 ） % and （70. 23 ± 1.36 ） %, Significantly lower than thecontrol group（P 〈0.01 ）. 3 ）In 10,12 mg/L group, there were statistically significant differences in the viability, most of cells deceased in the procedure of multi-directional differentiation. Conclusions PEI-SPIO of 8mg/L can effectively label hADSCs, without influencing the cell viability,proliferation and the multiple differentiation ability.