磁性二氧化硅纳米微球对人源脐带间充质干细胞活性的影响

Effects of magnetic silica nanospheres on the viability of human umbilical cord mesenchymal stem cells

目的探究磁性二氧化硅纳米微球作为一种干细胞表面的潜在标记物,对人脐带间充质干细胞(umbilical cord mesenchymal stem cells, UCMSCs)的表面干性标志物表达、细胞增殖和迁移能力等生物学活性的影响。方法先采用溶剂热法和St9ber方法制备一种生物相容性高的磁性二氧化硅纳米微球,再通过组织块培养法分离和原代培养UCMSCs,并依次采用显微镜、流式细胞术和三系分化等手段观察与鉴定其干细胞基本特性;其次,将不同质量浓度(1、10、50、100、200μg/mL)磁性二氧化硅纳米微球与上述细胞共孵育,并通过磁分离获得微球标记的干细胞;最后,通过倒置显微镜成像、流式细胞术、CCK-8法检测和细胞划痕实验等技术,评估其对干细胞的细胞形态、表面干性标志物表达、增殖和迁移能力等活性的影响。结果本研究成功分离和原代培养出了具有干细胞基本特性的UCMSCs;显微镜成像观察到不同质量浓度的磁性二氧化硅纳米微球均可标记UCMSCs;CCK-8法检测显示微球在浓度为1μg/mL时能够促进该干细胞的增殖,而随着微球质量浓度(10、50、100、200μg/mL)的增大,其对细胞增殖抑制作用亦增强;流式分析和细胞迁移结果表明10μg/mL微球对标记细胞的表面干细胞标记物表达和迁移能力无明显干扰。结论磁性二氧化硅纳米微球可用于标记UCMSCs,且在低浓度剂量下不会对该细胞表面干性标志物表达、增殖和迁移等活性造成显著干扰。

Objective To investigate the effect of magnetic silica nanospheres(MSNPs)on the viability of human umbilical cord mesenchymal stem cells(UCMSCs).Methods MSNPs with high biocompatibility were prepared with solvothermal method and Stober method.The UCMSCs were isolated and cultured with tissue block culture method.The basic characteristics of the stem cells were observed and identified with microscopy,flow cytometry and three-line differentiation.The UCMSCs were incubated with MSNPs at different concentrations(1,10,50,100,200μg/mL),and the MSNP-labeled stem cells were obtained by magnetic separation.The morphology,expression of stemness markers,proliferation and migration ability of MSNP-labeled stem cells were evaluated with fluorescence inverted microscopy,flow cytometry,CCK-8 detection,and cell scratch assay,respectively.Results The UCMSCs with the basic characteristics of stem cells were successfully isolated and cultured.Microscopy showed that UCMSCs were labeled by different mass concentrations of MSNPs.CCK-8 test showed that 1μg/mL microspheres promoted the proliferation of stem cells,while its inhibitory effect was enhanced with the increasing concentrations of MSNPs(10,50,100,200μg/mL).Flow cytometry and cell migration assay showed that 10μg/mL MSNPs did not significantly affect surface marker expression and migration of cells.Conclusion MSNPs can be used to label UCMSCs,without significant interfering with their active functions,such as stemness marker expression,cell proliferation and migration at low dose.