兔脂肪干细胞MR示踪前的超顺磁性氧化铁最佳标记条件筛选  

Screening of optimal labeling conditions of superparamagnetic iron oxide before MR tracing of rabbit adipose stem cells

作　　者：陈梦倩 罕迦尔别克·库锟 王云玲 贾琳 CHEN Mengqian;Hanjiaerbieke Kukun;WANG Yunling;JIA Lin(Department of Imaging Center,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China;Department of Medical Plastic and Cosmetic Surgery,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院影像中心,乌鲁木齐830054 [2]新疆医科大学第一附属医院医学整形美容中心,乌鲁木齐830054

出　　处：《新疆医科大学学报》2024年第7期938-944,共7页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金项目(2020D01C232)。

摘　　要：目的 采用多聚赖氨酸修饰的超顺磁性氧化铁(Polylysine coated superparamagnetic iron oxide,PLL-SPIO)纳米颗粒标记兔脂肪间充质干细胞(Adipose derived stem cell,ADSCs),探讨不同SPIO浓度及孵育时间对ADSCs标记率、存活率及细胞增殖率的影响,筛选SPIO标记ADSCs的最优标记条件。方法 用含不同浓度的SPIO(12、25、50μg/mL)培养基培养第3代兔ADSCs,分别孵育不同时间(6、12、24、48 h),孵育完成后,普鲁士蓝染色计算细胞标记率;细胞毒性实验检测细胞存活率;根据细胞毒性实验结果筛选细胞存活率较大的6组细胞进行细胞增殖能力检测,检测细胞增殖率。结果 普鲁士蓝染色结果显示,各组细胞标记率及细胞内蓝染颗粒数量,随着SPIO浓度增大及孵育时间的延长而增多。细胞毒性实验结果显示,在较低SPIO浓度和较短孵育时间下,细胞存活率不受影响,且各组之间细胞存活率无明显差异,随着SPIO浓度增大及孵育时间延长,细胞存活率呈下降趋势,48 h时细胞存活率下降最明显。细胞增殖实验结果显示,实验组各组细胞增殖率,随SPIO浓度增大及孵育时间延长而降低;且细胞进入快速增长期后,实验组细胞增殖率与阴性对照组相比,均存在明显差异,其中12μg/mL SPIO-6 h组和25μg/mL SPIO-6 h组细胞增殖率与阴性对照组差异较其他各组小。结论 从细胞内SPIO颗粒含量及细胞标记率、存活率及增殖率几方面综合评价,25μg/mL SPIO孵育6 h,是本研究所选出的PLL-SPIO标记ADSCs的最优标记条件。Objective To label rabbit adipose mesenchymal stem cells(Adipose derived stem cells,ADSCs) with polylysine modified superparamagnetic iron oxide(Polylysine coated superparamagnetic iron oxide,PLL-SPIO) nanoparticles,to explore the effects of different SPIO concentration and incubation time on ADSCs labeling rate,survival rate and cell proliferation rate,and to select the optimal labeling conditions for SPIO labeling ADSCs.Methods The 3^(rd) generation rabbit ADSCs was cultured with different concentrations of SPIO(12,25,50 μg/mL) and incubated for different time(6,12,24,48 h).After the incubation,prussian blue staining was used to calculate the cell labeling rate,cytotoxicity test was used to detect the cell survival rate,and 6 groups of cells with higher cell survival rate were selected according to the results of cytotoxicity test.Results Prussian blue staining showed that the cell labeling rate and the number of blue staining granules in each group was increased with the increase of SPIO concentration and the prolongation of incubation time.In the cytotoxicity test,the cell survival rate was not affected at lower SPIO concentration and shorter incubation time,and there was no significant difference among the 3 groups.With the increase of SPIO concentration and the prolongation of incubation time,the cell survival rate was decreased,and the decrease was the most obvious at 48 h.The results of cell proliferation experiments showed that the cell proliferation rates of the experimental groups decreased with the increase of SPIO concentration and the extension of incubation time;and after the cells entered the rapid growth period,there were significant differences in the cell proliferation rates of the experimental groups compared with those of the negative control group,among which the differences in cell proliferation rates of the 12 μg/mL SPIO-6 h group and the 25 μg/mL SPIO-6 h group and the negative control group were smaller than those of the other groups.Conclusion According to the comprehensive evaluatio

关 键 词：超顺磁性氧化铁 脂肪间充质干细胞 孵育浓度 孵育时间 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R445.2[医药卫生—基础医学]