CAJ | 학술논문

目的使用超小超顺磁氧化铁颗粒(USPIO)对大鼠脂肪源间充质干细胞(ADSCs)进行体外标记,研究不同浓度的USPIO对大鼠ADSCs生物学活性的影响,确定其标记ADSCs的适宜浓度. 方法实验分为8个组,其中6组依次添加不同终浓度的USPIO(180 μg/mL、135 μg/mL、90 μg/mL、45 μg/mL、22.5 μg/mL、11.25μg/mL),另2组为阴性转染组和空白对照组.普鲁士蓝染色检测USPIO标记ADSCs的效率,同时采用CCK-8及Alamar blue法检测各组细胞增殖情况. 结果 USPIO标记浓度在45 μg/mL时,普鲁士蓝染色后ADSCs胞浆内可见大量蓝色颗粒,标记效率在95％以上；USPIO标记浓度在90 μg/mL及以上时,标记效率约100％.CCK-8及Alamar blue检测结果表明USPIO在11.25～90 μg/mL范围内对细胞活力的影响较小且差异无统计学意义(P＞0.05),故可将45-90 μg/mL确定为适宜浓度. 结论 USPIO可在体外有效标记ADSCs,并且在适宜浓度范围内对细胞生物学活性无明显影响.
목적 사용초소초순자양화철과립(USPIO)대대서지방원간충질간세포(ADSCs)진행체외표기,연구불동농도적USPIO대대서ADSCs생물학활성적영향,학정기표기ADSCs적괄의농도. 방법 실험분위8개조,기중6조의차첨가불동종농도적USPIO(180 μg/mL、135 μg/mL、90 μg/mL、45 μg/mL、22.5 μg/mL、11.25μg/mL),령2조위음성전염조화공백대조조.보로사람염색검측USPIO표기ADSCs적효솔,동시채용CCK-8급Alamar blue법검측각조세포증식정황. 결과 USPIO표기농도재45 μg/mL시,보로사람염색후ADSCs포장내가견대량람색과립,표기효솔재95％이상；USPIO표기농도재90 μg/mL급이상시,표기효솔약100％.CCK-8급Alamar blue검측결과표명USPIO재11.25～90 μg/mL범위내대세포활력적영향교소차차이무통계학의의(P＞0.05),고가장45-90 μg/mL학정위괄의농도. 결론 USPIO가재체외유효표기ADSCs,병차재괄의농도범위내대세포생물학활성무명현영향.
Objective To label adipose-derived stromal cells (ADSCs) with different concentrations of ultrasmall superparamagnetic particles of iron oxide (USPIO) to investigate the biological characteristics of ADSCs treated with these USPIO,and determine the optimal concentration of USPIO in labeling ADSCs in vitro. Methods USPIO with different concentrations were prepared,and the particles were endocytosed by ADSCs generated from rat adipose tissue. Eight groups (negative control group,blank control group,and 11.25,22.5,45,90,135 and 180 μg/mL treatment groups) were chosen. Labeling efficiency and cellular uptake were analyzed by Prussian blue staining. Meanwhile,proliferation capacity and viability of ADSCs were evaluated by Alamar blue assay and Cell Counting kit-8. Results ADSCs could be effectively labeled with USPIO: approximately 95％ ADSCs were labeled when they were incubated with USPIO for 24 h under the concentration of USPIO was 45 μg/mL;and approximately 100％ ADSCs were labeled when the concentration of USPIO was 90 μg/mL and above. The CCK-8 and Alamar blue tests showed that USPIO of different concentrations (11.25-90 μg/mL) had little influence on cell growth viability,and no significant difference was noted between each 2 concentration groups (P＞0.05). In a word, 45-90 μg/mL USPIO were the optimal choice for transplantion of ADSCs in vivo. Conclusion ADSCs from the adipose tissue can be effectively labeled with USPIO with minimal effect on cell proliferation and viability.